House Fly Attractants and Arrestante: Screening

of Chemicals Possessing Cyanide, Thiocyanate,

or Isothiocyanate Radicals

Agriculture Handbook No. 403

Agricultural Research Service

UNITED STATES DEPARTMENT OF AGRICULTURE
Contents

Page

Methods 1

Results and discussion 3

Thiocyanic acid esters 8

Straight-chain nitriles 10

Propionitrile derivatives 10

Conclusions 24

Summary 25

Literature cited 26

This publication reports research involving pesticides. It does not contain recommendations for

their use, nor does it imply that the uses discussed here have been registered. All uses of pesticides

must be registered by appropriate State and Federal agencies before they can be recommended.

CAUTION: Pesticides can be injurious to humans, domestic animals, desirable plants, and fish

or other wildlife—if they are not handled or applied properly. Use all pesticides selectively and

carefully. Follow recommended practices for the disposal of surplus pesticides and pesticide

containers.

¿/áepé4áaUÁí^a¡eé

—' -"" TMK LABIL

Mention of a proprietary product in this publication does not constitute a guarantee or warranty

by the U.S. Department of Agriculture over other products not mentioned.

Washington, D.C. Issued July 1971

For sale by the Superintendent of Documents, U.S. Government Printing Office

Washington, D.C. 20402 - Price 25 cents
House Fly Attractants and Arrestants: Screening

of Chemicals Possessing Cyanide, Thiocyanate,

or Isothiocyanate Radicals

BY M. S. MAYER, Entomology Research Division,

Agricultural Research Service ^

Few chemicals possessing cyanide (-CN), thioeyanate (-SCN), or isothiocyanate (~NCS) radicals have been tested as attractants for the house

fly (Musca domestica L.). In fact, chemicals in

these groups apparently have received little consideration as attractants or arrestants for any

species. The distinction between attractant and

arrestant was carefully drawn by Dethier and

others (S).^ These authors pointed out that an

attractant is a chemical that causes insects to

make oriented movements toward its source, but

an arrestant is a chemical that causes insects to

aggregate in contact with it. Thus, an attractant

may work over a distance, but an arrestant must

be contacted by the insect to have an effect. Some

chemicals possessing these radicals as attractants

and arrestants have been reported (Beroza and

Green 1, Green and others 6, Jacobson 8) and

some of them have been reported as repellents.

Parman and others (12) found allyl isothiocyanate had little ability either to arrest or to

repel the screw-worm (Cochliomyia hominivorax

(Coquerel)). In a later publication the same group

(Laake and others 9) reported that allyl isothiocyanate was slightly attractive to Musca domestica, but it was considered to be one of the better

repellents for Phormia regina (Meigen). Gupta and

Thorsteinson (7) found allyl isothiocyanate to be a

weak attractant and arrestant for Plutella maculipennis (Curtis), and it was also reported (Dethier

2) to elicit a biting response by a mustard beetle,

Phaedon cochleariae (F.).

LaBrecque and Wilson (10) found benzhydryl

Ihiocyanate (diphenylmethyl thiocyanate) to be

ineffective as a repellent for house flies. Since in

this same report dioctylamine and two octyl sulfoxides were promising, Wilson and others (15)

tested a group of similar or related chemical repellents. Included in these tests were two nitriles,

4-(octylsulfinyl)butyronitrile and 4-(octylthio)-

butyronitrile, which appeared to be reasonably

good repellents. In my tests these two chemicals

failed to arrest a single fly. Ralston and Barrett

(IS) found that aliphatic nitriles containing 10 to

14 carbon atoms were more repellent to flies than

oil of citronella. In my tests no aliphatic nitriles

of these chain lengths were effective as arrestants.

METHODS

Two methods were used to screen the chemicals.

One was to assess the ability of the chemical to

attract house flies through olfactory stimulation

^ I thank the following people in this Division: M. W.

Abbott, G. S. Asbell, and J. D. James for screening the

chemicals, D. J. Guinyard for formulating most of the chemicals, Marie Osborne for assistance with the chemical nomenclature, and C. N. Smith for helpful criticism, guidance, and

encouragement.

2 Italic numbers in parentheses refer to Literature Cited,

p. 26.

with an olfactometer. The other was to assess the

ability of the chemical to arrest the flies through

odor, tarsal chemoreceptor contact, or both with

petri-dish tests.

The olfactometer technique has been described

by Mayer and Thaggard (11). By this technique

the flies were confined in the olfactometer, a large

cage with two circular ports at the front end. Each

port was fitted with a cylindrical trap closed by a

screen cover at the end away from the cage and a

screen funnel at the end toward the cage. Com-
AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

petitive attractants were exposed in pairs in

sleeves attached to the end of the traps farthest

from the cage. Air was drawn by a suction fan

through the sleeves, over the attractant, through

the trap, through the cage of flies, and then exhausted outside the building. After 30 minutes the

flies captured in the traps were counted and

returned to the cage.

The flies were from the regular laboratory

colony and were reared in the usual manner on

CSMA medium. All were reared under alternating

12-hour periods of light and darkness. When the

adult flies were 1 to 3 days old, they were counted

under carbon dioxide narcosis, placed in the olfactometer, and held under constant light. They were

not tested within 24 hours after narcosis. Water

was provided in the olfactometer at all times. The

food was the standard adult diet of powdered egg

yolk, milk solids, and sucrose, which was placed

in four small paper cups. For tests with ammonia,

the food remained in the olfactometer at all times.

For tests with the cyanide compounds, the food

was removed 2 hours prior to initial testing and

returned at the end of the day.

To begin each series of tests, 150 males and 150

females were placed in the olfactometer. During

the tests a few flies occasionally escaped. Dead

flies were counted daily and the number was subtracted from the computation of flies in the test,

but they were not removed from the cage. At the

end of each week, when the flies remaining in the

olfactometer were 7 to 10 days old and not suitable for further testing, they were killed and

counted. When this procedure was used by a capable technician, the final counts never showed

more than 8 percent error in the figures previously

obtained by subtraction.

The petri-dish technique (Dethier and others S)

was performed as follows: From 500 to 700 house

flies from the regular colony or an insecticideresistant strain (Cradson-P) were anesthetized

with carbon dioxide. Equal numbers of each sex

were placed in cylindrical screen holding cages

(20.3 cm. long and 6.3 cm. in diameter), provided

with a small amount of a 10 percent honey solution on absorbent cotton pads, and held overnight

in a room at 27° dz 1° C. and 50 ± 5 percent relative humidity. The next day, 18 hours after

anesthesia, the flies were released in the same room

into a cubical cage (1.22 meters on a side) and

allowed to feed on 10 to 20 grams of sugar for 30

minutes. During this interval all dead flies were

removed from the cage and counted so that the

number and sex ratio of those in the cage were

known. After the 30-minute feeding interval, the

sugar was removed; 30 minutes later it was offered

to the flies again and testing was begun only after

2 percent of the flies had begun to feed.

In several tests, flies held overnight in a relative

humidity of 70 ± 5 percent and tested at 50 percent relative humidity did not approach the food

as readily as those held at 50 ± 5 percent. Flies

held at the higher relative humidity tended to

show much less mortality than those held at the

lower humidity. These observations may indicate

that the flies at 70 percent relative humidity fed

better on the honey than at 50 percent. The percentage of the regular-colony flies that approached

the feeding dishes during testing was smaller than

that of the Cradson-P strain. Other observations

indicated that the percentage of flies feeding was

also affected by other factors such as their age and

overcrowding of the larvae in the rearing containers, as well as interactions between all these

factors. In all tests, however, the 2 percent feeding

criterion was strictly followed as the minimum

activity for beginning tests, and all chemicals were

retested if the number of flies on a sugar standard

fell below 2 percent of the total in the cage.

Solutions or suspensions of the test chemicals

in acetone or water were mixed thoroughly with

white fire-cleaned sand at the rate of 0.2 gram

of test chemical to 20 grams of sand. Two uncovered 9-cm. petri dishes, one containing the treated

sand and the other containing an equal volume of

granulated white sugar (about 13 grams) as a

standard, were exposed simultaneously in the cage

of flies. After 2 minutes the flies in each dish were

counted and disturbed from the dishes by a wave

of the hand, the positions of the dishes were reversed, and the test was repeated. Four 2-minute

exposure periods constituted a test. The number

of flies on the experimental material divided by

the number feeding on the standard (T/Std) gave

the ratio to the standard. This test was designed

primarily to select materials favored as foods, but

it may also show the presence of compounds

stimulating other responses as well as feeding.

Since these tests were performed with flies fed

within 2 hours prior to testing, I assume that the

foregut stretch receptor studied by Gelperin (4-, 5)

could have affected the threshold of response in
HOUSE FLY ATTRACTANTS AND ARRESTANTS

such a way as to lower the response of the flies, at

least in the petri-dish tests and possibly in the

olfactometer tests. However, in such a screening

program as this, a chemical that would still be

operative under these conditions would be even

better if the flies were starved.

Many of the chemicals used in these tests were

old and had been held in storage facilities subject

to the vagaries of outside temperature conditions.

In some instances, the chemical quite possibly had

become degraded. However, it was tested on the

assumption that all activity may not have been

lost. Most of the chemicals were crystalline, and

after formulation they were always held under

refrigeration.

RESULTS AND DISCUSSION

For many years olfactory attractants specific

for the house fly have been sought by screening or

testing in glass traps similar to those described by

Parman and others (12) and LaBrecque and Wilson (10), Generally this technique has not been as

satisfactory as desired because of its extreme variability. For this and other reasons, the olfactometer

technique as described here was developed for

testing attractants where olfactory stimulation

must be followed by positive anemotactic or

klinokinetic movement.

Edamin, a hydrolyzed milk protein solid, was

used as the standard, although it has only slight

capacity for olfactory stimulation. Where proteinaceous baits such as Edamin are subjected to bacterial decomposition, ammonia or other bacterial

metabolic byproducts or both probably account

for most of the attractiveness. The results of

preliminary tests with Edamin, water, and

ammonium hydroxide are shown in table 1. The

tests were paired comparisons and analyzed

accordingly. Edamin was considered a good standard for comparison because (1) it is slightly more

attractive than water; (2) the standard errors of

the means were low; (3) little sexual bias existed;

(4) the presence or absence of food in the cages

failed to produce a marked difference in the response; and (5) Edamin rarely failed to attract

some flies during these tests. When Edamin was

compared with 1.0 N ammonium hydroxide, the

males definitely preferred Edamin, whereas the

females, as expected, preferred 1.0 N ammonium

hydroxide.

TABLE 1.—Attractiveness of Edamin, water, and ammonium hydroxide to house

fies with and without food in olfactometer tests

Attractant ^ Males trapped ^ Females trapped '

Replications

(number) Port 1 Port 2 Port 1 Port 2 Port 1 Port 2

Percent Percent Percent Percent

WITH FOOD IN CAGE

7 Dry Edamin Wet Edamin.

.... do Water

Wet Edamin do

11

WITHOUT FOOD IN CAGE

pry Edamin Wet Edamin 7.3

6 \ do Water

[Wet Edamin do

12 Wet Edamin 1.0 N ammonium

hydroxide.

8.1 8.5 5.5 3.6

8.6 3.6 8.7 2.1

10.4** 4.9 5.0 3.5

CAGE

7.3 6.7 4.1 7.8

9.0 5.5 3.4 3.6

5.9 4.9 8.2* 3.6

8.3* 4.4 6.0 14.6^

^ Wet Edamin = 1 gram of Edamin with enough water (about 5 ml.) to give consistency of

batter.

2 Statistical significance in paired comparisons indicated by asterisks (* = 0.05 level, ** =

0.01 level).
AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

Since considerable data existed on the attraction

of house flies to ammonia (Richardson lU^ and

others), I used ammonia to test the olfactometers,

techniques, and flies to determine whether the

percent attraction conformed to a logarithmic

increase relative to the concentration used.

Ammonia was introduced into the cages by placing

2 ml. of solution containing various concentrations

of ammonium hydroxide on filter paper in the

bottom of Syracuse watch glasses. The solutions

were replaced at 10-minute intervals during the

30-minute test period.

The results of tests in which ammonia was introduced in an ascending order of concentration

are presented in figure 1. This graph is a compilation of data for flies of several generations and

ages. The concentration-response relationship conforms to a straight line over four log units of concentration, with the response decreasing at the two

highest concentrations. The method employed for

introducing the ammonia is primitive and was

meant to test the flies under conditions intended

for screening other chemicals. The standard errors

were large. However, within most groups of flies

of the same generation the line appeared straight

over six log units and would then, of course, conz

o

20 1 1 1

/-\

15-

/

/

\

10- /

<

fi

> 0 ^ E« .«>H Bl^»a

O

MH ^m ■■ n

o

0 - 1 \ 1

2

log C

FIGURE 1.—Attraction of 5- to 7-day-old female house flies

to various concentrations of ammonium hydroxide when

offered in ascending order (solid line) as compared to

male and female response to water and male response to

ammonium hydroxide (control) (dotted hne).

form to the Weber-Fechner relationship over the

entire range tested.

A true sensory response is evident in the data

obtained when the concentrations were tested in

descending order, as shown in figure 2. The highest

concentration induced a higher response when

offered at the beginning of the series than when

offered after the lower concentrations (fig. 1). This

was followed by an adaptation to all lower concentrations and hence a reduced response. In fact, the

concentration that gave the highest response when

offered in ascending order produced a response

in this series that was not statistically above the

threshold (about 5 percent). For these reasons, the

olfactometers were presumed to measure accurately the olfactory stimulation of various chemicals that produce behavioral change. Additional

data showed no response by females prior to 5 days

of age and no response after oviposition, indicating that the olfactory response to ammonia is

dependent on ovarian development.

A screening program of potential olfactory

attractants for the house fly was undertaken with

these olfactometers. Early in this program certain

nitriles were observed to stimulate a large number

of females to enter the paired control port baited

with Edamin. The results of olfactory tests with

FIGURE 2.—Attraction of 5- to 7-day-old female house flies to

various concentrations of ammonium hydroxide when

offered in descending order, based on six to eight replications for each point.
HOUSE FLY ATTRACTANTS AND ARRESTÁNTS

0 CH2CH2CN

/ /

C—N

r-s] CH2CH2CN

\J cm

ENT-26949

52 nitriles are presented in table 2. None of the

compounds were highly attractive, but several

were associated with unusually high response to

Edamin. Two possible behavioral responses could

produce these results: (1) The entry of the flies

into the Edamin trap could be an avoidance

response to the nitrile (repellency), since the

Edamin trap was the only part of the olfactometer

accessible to the flies that was free of the nitrile

odor or (2) the nitrile odor could stimulate a feeding response or a search for food by the females,

which then orient to the odor of Edamin. Later

tests with the petri-dish technique were to give

some indication as to whether the compounds were

actually repellents or feeding stimulants.

The first nitrile derivative screened in petri

dishes was iV,A/'-bis(2-cyanoethyl)-m-toluamide

(ENT-26949). A high positive response

(ratio T/Std = 5.3:1) to this chemical was obTABLE 2.—Attractiveness to house flies of Edamin and various nitriles, listed in order of female response

to Edamin, in paired olfactometer tests (S replications)

served (table 3). The attractiveness or arrestancy

of this chemical was apparently due to the

presence of the cyano groups, since AT^iV-diethylm-toluamide (ENT-20219), which is otherwise

0

/ /

C—N

CH2CH3

O

CH2CH3

CHs

ENT-20219

Code No.

(BNT-)

Chemical

Attractancy of

Edamin to—

Attractancy of

chemical to—

Males Females Males Females

Percent Percent Percent Percent

16234 l-Naphthalenepropionitrile, 2-hydroxy16310. Benzonitrile, m-mtro17792 Propionitrile, 3-(??-chlorobenzoyl)-2-[3,4-(methylenedioxy)phenyl]-

17128 o-Tolunitrile

17129 m-Tolunitrile

17436 Acetonitrile, diphenyl18677 Heptanedinitrile, 4-acetyl-4-phenyl14826 p-Tolunitrile

16034...... Heptanedinitrile, 4-(??-bromobenzoyl)-4-(2-cyanoethyl)-

28304.. 2-Decenenitrile

13188 4-Morpholinepropionitrile

310 Myristonitrile

16027 Heptanedinitrile, 4-benzoyl-4-(2-cyanoethyl)-

28305 2-Nonenenitrile

14265 4-Morpholinesuccinonitrile ,

20145 Mandelonitrile, 3,4-(methylenedioxy)-

49 Phthalonitrile

4979 1-Cyclohexene-l-acetonitrile

17751 Glycinonitrile, N-methylene16033 Heptanedinitrile, 4-(2-cyanoethyl)-4-2)-toluyl16026 1,1,3,3-Cyclopentanetetrapropionitrile, 2-oxo17359 Acetonitrile, bis[p-(dimethylamino)phenyllphenyl7618 Heptadecanenitrile

8103 Sebaconitrile

4981 1-Cyclohexene-l-acetonitrile, 2-allyl234 2-Naphthonitrile

15766 Nicotinonitrile

9.1 42.8 6.8 2.5

6.0 37.4 6.6 2.5

8.4 27.5 7.1 2.4

7.6 26.7 3.6 .9

7.8 25.5 8.6 2.1

11.1 25.0 8.1 2.1

7.5 25.0 9.7 2.0

12.9 23.7 4.6 2.8

4.8 23.6 8.2 4.8

4.0 22.6 2.6 .2

8.7 21.6 5.9 4.4

17.3 16.6 10.6 3.7

5,3 16.1 6.4 5.4

7.3 14.6 1.1 3.5

13.3 14.4 10.1 4.0

3.0 13.8 8.2 2.5

12.1 13.0 6.9 1.4

5,2 12.3 .9 .5

6.6 12.1 9.6 3.1

4.3 11.4 5.3 13.1

4.9 11.4 4.4 1.9

6.7 11.1 6.5 2.0

10.4 10.9 9.4 3.3

9.1 10.9 8.9 5.8

7.6 9.6 2.4 3.1

6.4 9.4 7.2 3.5

4.0 9.3 6.1 3.4
AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

TABLE 2.—Attractiveness to house flies of Edamin and various nitriles, listed in order of female response

to Edamin, in paired olfactometer tests (3 replications)—Continued

Code No.

(ENT-)

Chemical name

Attractancy of

Edamin to—

Attractancy of

chemical to—

Males Females Males Females

Percent Percent Percent

17661 Cyclopropanecarbonitrile, 2-benzoyl-l,3-diphenyl478...... Benzonitrile, p-nitro18124 Cyclohexanecarbonitrile, 1-heptyl1547 Nicotinonitrile, l,2-dihydro-l,4,6-trimethyl-2-oxo16570 Tridecanenitrile

15707 Lactonitrile, acetate

16030..... . Heptanedinitrile, 4-(p-chlorobenzoyl)-4-(2-cyanoethyl)- . . . .

13063 Heptanedinitrile, 4-acetyl-4-methyl11234... . . .Palmitonitrile.

7106 Hexanenitrile, 2,4-dihydroxy-3,3,5-trimethyl15930 Butyronitrile, 4-benzoyl-2-(dp/ia-phenacylbenzyl)-2,3-diphenyl-. .

16089 Heptanedinitrile, 4-(2-cyanoethyl)-4-(3,4-dichlorobenzoyl)-—.. .

16090 Heptanedinitrile, 4-benzoyl-4-methyl4364 Acetonitrile, benzoyl54......Acrylonitrile

11298..... .Benzonitrile, 2,4,6-triisopropyl11801 p-Anisonitrile

8777 Propionitrile. .

4975 Acetonitrile, phenyl11080... . . .Adiponitrile

311 Lauronitrile

28307 2-Furanacrylonitrile

1651..... . 3-Cyclohexene-l-carbonitrile, 5,5-dimethyl-2-(l-methylpropenyl)-.

28306 2-Octenenitrile, 5,7,7-trimethyl- ..

7024 Glutaronitrile

7.9

4.5

8.2

8.1

7.6

5.9

8.7

9.2

11.8

9.3

4.9

3.7

6.9

10.3

6.8

8.8

7.1

8.1

9.1

8.7

7.3

9.2

4.5

5.8

7.5

8.8

8.8

8.2

7.4

7.4

7.1

6.9

6.8

6.6

6.3

6.0

5.8

5.7

5.7

5.3

4.7

4.5

4.4

4.3

4.2

3.9

3.6

2.9

2.5

13.0

5.9

5.

5.

4.

5.

6.6

7.9

9.2

8.6

4.7

9.8

6.7

13.8

5.4

8.3

7.5

8.8

4.8

7.5

7.0

3.4

3.3

3.8

7.7

Percent

5.1

1.1

1.3

4.7

3.3

4.9

4.9

4.9

5.6

4.3

2.4

2.3

.6

4.0

2.2

3.3

2.1

4.5

2.4

2.1

3.5

1.0

1.7

1.6

7.4

structurally identical, is one of the best insect

repellents. However, I also considered the cyanoethyl radical (I) or propionitrile (ENT-8777) as

failed to arrest any flies. The chemicals were

classified as follows according to their arrestancy

to flies:

-CH2CH2CN

I

CH3CH2CN

ENT-8777

having a potential worth investigating. Thus,

early in this program certain known repellents and

derivatives were screened to eliminate any likelihood that toluamide and benzamide derivatives

were attractive.

The ratios of the numbers of flies on the treated

sand to the numbers on granulated sugar were

computed (table 3). In addition, from the results

with all chemicals tested, five class groupings were

assigned, based on the total number of flies

arrested on the treated sand, regardless of the

numbers found on the sugar standard. This

arrangement is highly artificial and not conducive

to analytical calculations; however, it was adopted

for ease of visualizing all the data. Untreated sand

Class

1

2

3

4

5

Flies (number) arrested

by chemical

0

1-4

5-25

26-100

Over 100

The toluamides and related compounds that

showed some arrestancy are listed in table 3. The

completely ineffective compounds were as follows:

Code No.

(ENT-) Chemical

26772 m-Toluamide, ATjiV-dicyclohexyl26774 m-Toluamide

30105. ... .m-Toluamide, iV,iV-dimethyI30033 o-Toluamide, Ar,Ar-dimethyl30330. . . . .2?-Toluamide, Ar,iV-dimethyl-
HOUSE FLY ATTRACTANTS AND ARRESTANTS

Code No.

iENT-)~Con. Chemical

20219 2}-Toluamide, 2V,Ar-diethyl20218 w-Toluamide, N^AT-diethyl20217 o-Toluamide, Ar,N-diethyl26779 m-Toluamide, N,N-diethy\-alpha,alpha,alpha'

trifluoro26781 m-Toluamide, NfN-diethyl-alpha-hydroxy30126 m-Toluamide, Ar,iV-dibutyl30002 o-Toluamide, N,Ar-dibutyl30146 p-Toluamide, Ar,2V-dibutyl32948 m-Toluamide, AT-ethyl-iV-isobutyl32952 m-Toluamide, iV-ethyl-N-propyl26059 m-Toluamide, AT-ethoxy-N-ethyl30355 p-Toluamide, AT-butyl32943 p-Toluamide, AT-butyl-iV-methyl32950 m-Toluamide, AT-butyl-AT-methyl20931 m-Toluamide, AT-isobutyl32953 m-Toluamide, N-isobutyl-N-propyl19775 Benzamide, A/'jAT-dibenzyl-p-ethoxy19084 Benzamide, ATjAT-dibutyl-o-ethoxy19710 Benzamide, A/',Ar-dibutyl-p-ethoxy20297 Benzamide, o-ethoxy-ATjiV-diethyl19708 Benzamide, AT-butyl-p-ethoxy20701 Benzamide, m-chloro-iV,iV-diethyl17586 Benzamide, o-chloro-iVjiV-diethyl20698 Benzamide, A/',Ar-diethyl-3,5-dimethyl20490 Benzamide, iVjAT-diethyl-o-isopropoxy20454 Benzamide, ATjN-diethyl-o-propoxy20690 Benzamide, A^,Ar,o-triethyl22386 Benzamide, Ar,N-dibutyl-o-chloro30182 Benzamide, ATjAT-dibutyl-p-isopropyl1016 Benzamide, A/'-pentyl19715 Benzamide, jj-ethoxy-AT-pentyl19716 Benzamide, p-ethoxy-ATjiV-dipentyl16750 Benzamide, AT-cyclohexyl-iV-pentyl20596 Phthalamide, iV,Ar,Ar',iV'-tetraethyl17014 p-Anisamide, iV,iV-diisopropylNone of the compounds were in class 5 except A/',A/^-bis(2-cyanoethyl)-m-toluamide (ENT26949), and only two were in class 4. In addition,

ENT-26949 was the only compound with a ratio

to sugar in excess of 1.0. Thus, it was reasonable to

assume that the cyano or the cyanoethyl moiety

was the principal causative agent of the arrestant

activity exhibited by ENT-26949. Therefore, a

large-scale screening program was accomplished

with its ultimate aim to determine which possible

combinations of structures carrying the cyanide

(-CN), thiocyanate (-SCN), and isothiocyanate

(-NCS) radicals could be responsible for the

arrestancy of house flies. To date, 390 chemicals

have been screened in this program.

The results of this system of classification for all

chemicals tested except those in table 3 are presented in figure 3. This graph illustrates several

TABLE 3.—Arrestant activity of various toluamides,

benzamides, and other compounds in petri-dish

tests

Code No.

(ENT-) Chemical

Ratio

Class T/Std

26949 m-Toluamide, N.N-h'm-

(2-cyanoethyl) 5 5.3:1

26773 m-Toluamide, N,iV-dibenzyl- . . 4 i .431:1

31518 m-Toluamide, iV,Ar-dioctyl- .... 4 .2:1

26950 m-Toluamide, iV,2V-diethylthio- 3 i .205:1

20930 o-Toluamide, AT-isobutyl- 3 .03:1

30079 o-Toluamide, 2V-benzyl- 2 i .05:1

26804 m-Toluamide, N,N-di'2-

pyridyl- 2 .02:1

32949 m-Toluamide, AT-butyl-ATethyl- 2 .01:1

20946 p-Toluamide, N-isobutyl- 2 .01:1

20680 Benzamide, p-tert-hutyl-N.Ndiisopropyl- 2 .03:1

19714 Benzamide, ZV-benzyl-p-ethoxy- 2 .02:1

19204 Benzamide, Ar,N-dibenzyl-oethoxy- 2 .02:1

28462 m-Toluidine, iV,Ar-diethyl- ..... 3 .08:1

1 Two or more tests.

features of the power of arrestancy for this entire

group of chemicals. No thiocyanates or isothiocyanates were found that could be included in

classes 4 and 5. However, only 61 thiocyanates

were tested. If the probability of finding a class

I

CLASS

FIGURE 3.—Proportion of all cyanides (-CN) and thiocyanates (-SCN) as arrestants in each class, based on

petri-dish tests.
8 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

4 or 5 arrestant was the same as for the cyanides,

then only two class 5 arrestants could have been

expected and four in class 4. Thus, with only six

chemicals expected to be class 4 or 5 arrestants,

the probability of finding them with this number

of chemicals tested may not be too good. Of course,

it is possible that no thiocyanate will be a class 4 or

5 arrestant.

Of all cyanides tested, 39.8 percent were considered active; 17.7 percent were in class 2, 10.2

percent in class 3, 5.9 percent in class 4, and 3.7

percent in class 5.

Prior to discussing the organic cyanides, the

results with three inorganic cyanides should be

noted. Although none of the three were outstanding arrestants, sodium pentacyanonitrosylferrate(/J/) (sodium nitroferricyanide) (ENT28742) was a class 3 arrestant (ratio 0.25), and

even cupric cyanide (ENT-904) was slightly

active (ratio 0.01). Sodium cyanide was inactive.

Thiocyanic Acid Esters

Results with various types of thiocyanates that

showed some arrestancy are given in table 4.

None of the aliphatic esters of thiocyanic acid

approached the magnitude of activity shown by

certain of the straight-chain cyanides (table 6).

Some small activity was observed in those with

2, 16, and 18 carbon chains. The ineffective

compounds were as follows:

Code No,

(ENT-) Chemical

16178 Thiocyanic acid, methyl ester

18432 Thiocyanic acid, heptyl ester

1125 Thiocyanic acid, octyl ester

1127. . . . .Thiocyanic acid, decyl ester

114 Thiocyanic acid, dodecyl ester

1128 Thiocyanic acid, tetradecyl ester

Virtually all of a group of alkyl p-hydroxyphenyl thiocyanates were at least slightly active;

only 5-¿er¿-butyl-4-hydroxy-m-tolyl thiocyanate

(ENT-19643) was inactive. In certain instances,

discussed later (p. 14), hydroxyl groups may have

induced great activity in a few chemicals where

such a degree of arrestancy was not expected.

Slightly more than half of a group of complex

thiocyanates showed some arrestancy. The

ineffective compounds were as follows:

Code No.

(ENT-) Chemical

15788 Nicotine, compound with cadmium thiocyanate

and thiocyanic acid (2:1:2)

17355 Pyrrolidinium, l-methyl-l-octyl-2-

{3-pyridyl).... thiocyanate

15825 Pyrrolidinium, l-butyl-l-methyl-2-

(3-pyridyl).... thiocyanate

15804 Pyrrolidinium, l-hexadecyl-l-methyl-2-

(3-pyridyl).... thiocyanate

17354 Pyrrolidinium, l-benzyl-l-methyl-2-

(3-pyridyl) thiocyanate

17353 Pyrrolidinium, l-(o-chlorobenzyl)-lmethyl-2-(3-pyridyl). . , .thiocyanate

1412 Nicotine, compound with cuprous thiocyanate

(2:1)

A few miscellaneous thiocyanates that showed

some activity are listed in table 4. Miscellaneous

ineffective compounds were as follows:

Code No,

(ENT-) Chemical

8887 Thiocyanic acid, benzyl ester

21778 Thiocyanic acid, phenacyl ester

7797 Thiocyanic acid, benzyl pentyl ester

14724. . . . .Propionic acid, 3-thiocyanato-, 6-propylpiperonyl ester

14725 Valeric acid, 2-thiocyanato-, 6-propylpiperonyl

ester

17008 Thiocyanic acid, pentachlorocyclohexyl ester

19643 Thiocyanic acid, S-¿er¿-butyl-4-hydroxy-m-tolyl

ester

4258-X.. .Thiocyanic acid, mixed esters with 1-chloro3-(isobornyloxy)-l ,2-propanediol

25370. . . . .Acetic acid, thiocyanato-, 2-(isobornyloxy)-

ethyl ester

19555 Crotonic acid, 3-thiocyanatopropyl ester

4789 Propionic acid, 2-thiocyanato-, ester with

dipentenecarbinol

4788 Propionic acid, 2-thiocyanato-, ester with

dipentenecarbinol, hydrogenated

2567.... .Thiocyanic acid, diester with triethylene glycol

3999 Thiocyanic acid, diester with diethylene glycol

(Lethane A-70)

8 .... . Lethane 384 Special (aliphatic thiocyanate)

5 Laurie acid, 2-thiocyanatoethyl ester (Lethane

60)

7 Lethane 384 Special; 50 percent mixture of three

parts 2-thiocyanatoethyl esters of aliphatic

acids and one part 2-(2-butoxyethoxy) ethyl

thiocyanate

Some of these are insecticides (Lethane series).

Only eight isothiocyanates were screened (table

5), Three of these demonstrated some activity, but

none were outstanding.
HOUSE FLY ATTRÁCTANTS AND ARRESTANTS

TABLE 4.—Arrestant activity of various thiocyanates in petri-dish tests

Code No.

(ENT-) Chemical Class

Ratio

T/Std

ALIPHATIC THIOCYANATES

18429 Thiocyanic acid, ethyl ester

1126 Thiocyanic acid, hexadecyl ester.

1124 Thiocyanic acid, octadecyl ester.

ALKYL p-HYDROXYPHENYL THIOCYANATES

19635 Thiocyanic acid, 4-hydroxy-m-tolyl ester

19636 Thiocyanic acid, 4-hydroxy-o-tolyl ester

19638..... .Thiocyanic acid, 4-hydroxy-2,6-xylyl ester

19639 Thiocyanic acid, 4-hydroxy-3,5-xylyl ester

19637 Thiocyanic acid, 3-ethyl-4-hydroxyphenyl ester.

19641 Thiocyanic acid, 4-hydroxy-w-cumenyl ester..,

19642 Thiocyanic acid, 5-hydroxycarvacryl ester

COMPLEX THIOCYANATES

15792 Nicotine, compound with cobaltic thiocyanate and thiocyanic acid (2:1:2). . . .

15780 Nicotine, compound with manganous thiocyanate and thiocyanic acid (2:1:2).

15778 Nicotine, compound with cuprous thiocyanate and thiocyanic acid (1:1:1)....

16795 Nicotine, compound with cuprous cyanide and thiocyanic acid (1:1:1)

1415 Nicotine, compound with cupric thiocyanate (1:1)

15784 Nicotine, compound with cupric thiocyanate and thiocyanic acid (2:1:2)

15837 Nicotinium compound, N,A?''-didodecyl-dithiocyanate

15823 Pyrrolidinium, l-methyl-l-(??-nitrobenzyl)-2-(3-pyridyl) thiocyanate

MISCELLANEOUS THIOCYANATES

14723. Acetic acid, thiocyanato-, 6-propylpiperonyl ester..

16979 Thiophene, 5-chloro-2-(l,2-dithiocyanatoethyl)-. . .

188 Thiocyanic acid, 4-methyl-l-naphthylmethyl ester.

4784 Thiocyanic acid, tris(p-chlorophenyl)methyl ester..

18233 Thiocyanic acid, diester with p,î>Mminodiphenol...

22371 Propionic acid, 2-thiocyanato-, ethyl ester

2 12 0.085

2 .013

2 .033

3 1 .12

2 .03

3 .31

2 1 .05

3 .43

2 1 .01

2 2 .15

3 1 .40

2 .09

2 2 .08

2 2 .07

3 .07

2 12 .03

2 2 ,02

2 .01

2 1 .01

2 2 .07

2 1 .11

2 .03

3 .12

2 .02

Seemed to stimulate flies more than ratio would indicate.

Two or more tests.

TABLE 5.—Arrestant activity of various isothiocyanates in petri-dish tests

Code No.

(ENT-) Chemical

Ratio

Class T/Std

16038. Isothiocyanic acid, 4-biphenylyl ester

16158 Isothiocyanic acid, 2?-(2-pyrimidylsulfamoyl)phenyl ester.

23336 . .Isothiocyanic acid, 1,1,3,3-tetramethylbutyl ester

15283 .Isothiocyanic acid, allyl ester

16039 Isothiocyanic acid, 2?-bromophenyl ester

16040 .Isothiocyanic acid, p-chlorophenyl ester

17006 .Isothiocyanic acid, 3-methoxy-l-vinylpropyl ester

17005 Isothiocyanic acid, 3-butoxy-l-vinylpropyl ester

3 0.09

3 .23

2 .05

0

0

0

0

0
10 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

In consideration of the relatively small number

of thiocyanates and isothiocyanates tested for

house fly arrestancy, no final conclusion should be

drawn of the potential activity of this group of

chemicals. However, it does not seem likely that

any will compare favorably with the nitriles in

activity. However, the thiocyanic acid radical

should be synthesized onto the same molecular

structures as other class 5 cyanides for a true

comparison of activity.

Straight-Chain Nitriles

Twelve nitriles were tested that differed only in

the length of the hydrocarbon chain (table 6).

None with chains more than five carbon atoms

long exhibited any activity, and valeronitrile

(ENT-30052) demonstrated only slight activity.

CH3(CH2)3CN

ENT-30052

Propionitrile (ENT-8777) was the best of the

short-chain nitriles, whereas acetonitrile (ENT327) and butyronitrile (ENT-8778) were slightly

CH3CH2CN CH3CN CH3(CH2)2CN

ENT-8777 ENT-327 ENT-8778

less effective. Isobutyronitrile (ENT-28525) (not a

straight-chain compound, see p. 16) was also

about equal to acetonitrile and butyronitrile.

CH2CH2CN

H2C

HsC

H3C

CHCN

ENT-28525

Since these straight-chain nitriles were active,

the next logical step was to test derivatives with

two cyano groups. The results with this group are

also given in table 6. None demonstrated activity

greater than class 3.

Since the straight-chain mononitriles that were

active had boiling points below 141° C, it is possible that this physical parameter is important for

activity. Although pimelonitrile (ENT-28713) was

not active, two of its derivatives (table 14) were

class 4 arrestants. This structure could be considered as two propionitriles if the fourth carbon

atom is considered a common substitution to both

chains.

CH2CH2CN

ENT-28713

Propionitrile Derivatives

The simplest unsaturated nitrile is acrylonitrile

(ENT-54). Although this compound was in class 4

(ratio 0.09), its activity never approached that of

propionitrile. A phenyl group on the 2 position

(ENT-7248) reduced the activity only slightly

(class 3, ratio 0.08), whereas a furan ring (ENT28307) on the 3 position abolished all activity.

An acetylenic bond (ENT-11802) also removed all

activity.

TABLE 6.—Arrestant activity of various straightchain aliphatic nitriles and dinitriles in petridish tests

Boil- CarCode No. ing bon Ratio

(ENT-) Chemical point atoms Class T/Std

° C. Number

NITRILES

. Acetonitrile 81 2

.Propionitrile 97 3

. Butyronitrile 118 4

. Valeronitrile 141 5

. Hexanenitrile 162 6

. Heptanenitrile 183 7

. Decanenitrile 244 10

. Lauronitrile 198 12

. Tridecanenitrile 13

. Myristonitrile 226.5 14

. Paimitonitrile 251.5 16

. Heptadecanenitrile 17

DINITRILES

. Malononitrile 220 3

. Succinonitrile 267 4

. Glutaronitrile 286 5

.Adiponitrile 295 6

-Pimelonitrile 169 7

. Suberonitrile 185 8

. Sebaconitrile 10

327.

8777.

8778.

30052.

28396.

28301.

11101.

311.

16570.

310.

11234.

7618.

24285.

6591.

7024.

11080.

28713.

33237.

8103.

5 0.80

5 2.64

5 1.99

2 .02

0

0

0

0

0

0

0

0

2 1 .04

1 10

3 .17

1 0

1 10

1 10

1 0

1 Two or more tests.
HOUSE FLY ATTRACTANTS AND ARRESTANTS 11

H2C = CHCN

ENT-54

CH = CHCN

CH2 C-

/\

0

-CN

ENT-7248

/X C = CCN

0

v

ENT-11802

\n/

ENT-28307

Most substitutions on the 3 position of

propionitrile resulted in a decrease or complete

loss of activity (table 7). However, certain relationships can be observed that will be important

in further work. Substitution of rings and substitution through ether, ester, and amino linkages

generally resulted in virtually total loss of activity.

However, substitution through sulfur linkage did

not affect the activity as much as the other types

and groups. The flies definitely preferred 3-(otolylthio)propionitrile (ENT-16985) over the meta

isomer (ENT-16986).

SCH2CH2CN

/\CH3

O

ENT-16985

Substitution of meta- or para-chlorophenyl

groups may also enhance activity. It is observed

that 3- (p-chlorophenoxy)propionitrile (ENT16700) was the only active ether, and 3-[(p-chlorophenyl)thio]propionitrile (ENT-16966) was the

best of the thio derivatives. The hydracrylonitrile

ester of m-chlorocarbanilic acid (ENT-24084) was

slightly active. Generally it seems that monosubOCH2CH2CN

O

SCH2CH2CN

o

Cl

ENT-16700

Cl

ENT-16966

H O

I r

/\NC—OCH2CH2CN

O

\/ci

ENT-24084

stitutions on the 2 position of propionitrile will

decrease activity unless such substitutions occur

through sulfur atoms.

The following chemicals with single additions

to a propionitrile molecule on the 2 position were

inactive:

Code No,

{ENT-) Chemical

18990 Valeronitrile, 2-methyl-3-oxo7461 Propionic acid, 2-cyano-, 4-methylcyclohexyl

ester

12066 3-Hexenal, 2-(2-cyanoethyl)-2-ethyl15707 Lactonitrile, acetate

23081 Carbanilie acid, m-chloro-, ester with lactonitrile

7470b.... Propionic acid, 2-cyano, 2-methylcyclohexyl

ester

26107 Nonanoic acid, 2-cyanoethyl ester

All but two of these were linked by an ester,

which will be shown later to interfere with activity.

However, 2-methyl-3-oxo-valeronitrile (ENT18990), which might be considered as a two-substituted propionitrile, showed some activity (class

2, ratio 0.02). Since no other chemicals with two

0 CH3

\ I

H3CCH2CCH

I

CN

ENT-18990

substitutions on propionitrile were available, it

cannot be ascertained whether all such substitutions will adversely affect activity.

Table 8 presents the results with some active

compounds, which are essentially di-substituted

propionitrile derivatives. The inactive compounds

were as follows:

Code No.

(ENT-) Chemical

50880 2-Aziridinecarbonitrile, 1-butyl23854 Hydratroponitrile, 6eia-(cyclohexylamino)-,

hydrochloride

4924 1-Cyclohexene-l-acetic acid, alpha-cya,no-alphamethyl-, ethyl ester

17203 2-Furanacrylonitrile, a^p/^a-phenyl5830 Hydrocinnamic acid, alpha-cyano-alpha-phenyl-,

ethyl ester

Two substitutions on the basic propionitrile

molecule superficially may appear to provide more

active compounds than only one substitution.
12 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

TABLE 7.—Arrestant activity of propionitrile derivatives substituted on 3 position in petri-dish tests

Code No.

(ENT-) Chemical Class

Ratio

T/Std

SINGLE INORGANIC SUBSTITUTION

17851 Propionitrile, 3-bromo- 0.07

SINGLE ORGANIC SUBSTITUTION

5926 .Butyraldehyde, 2-(2-cyanoethyl)-2-ethyl-.

RING SUBSTITUTION

23733 Hydrocinnamonitrile

14830 Cyclohexanepropionitrile, 1-nitro18069 Propionitrile, 3-isobornyl13188 4-Morpholinepropionitrile

17124 L-Pyroglutamic acid, l-(2-cyanoethyl)-. .08

SUBSTITUTION THROUGH ETHER LINKAGE

25449 Propionitrile, 3-methoxy25450 .Propionitrile, 3-ethoxy24894 Propionitrile, 3,3'-oxydi-.

13078 Propionitrile, 3-(allyloxy)

17772 Propionitrile, 3-isopropoxy8628 — . . Propionitrile, 3-butoxy17914 Propionitrile, (hexyloxy)-

8290 Propionitrile, 3-[(2-ethylhexyl)oxy]-

5764 .Propionitrile, 3-(cyclohexyloxy)-

5935 Propionitrile, 3-[(2-methylcyclohexyl)oxy]-.

16700 .Propionitrile, 3-(??-chlorophenoxy)-

14827 Propionitrile, 3-(2-phenoxyethoxy)-

0

0

10

10

0

10

0

10

0

10

.55

SUBSTITUTION THROUGH SULFUR LINKAGE

14259 Propionitrile, 3-(phenyIsulfonyl)-

16966..... .Propionitrile, 3-[(p-chlorophenyl)thioJ-.

16986 Propionitrile, 3-(m-tolylthio)-

16985 .Propionitrile, 3-(o-tolylthio)-.

16840.. Propionitrile, 3,3'-thiodi-

.12

1.05

0

.13

0

SUBSTITUTION THROUGH ESTER LINKAGE

2885 Acrylic acid, 2-cyanoethyl ester

16824 Cinnamic acid, 2-cyanoethyl ester

24084 Carbanihc acid, m-chloro-, ester with hydracrylonitrile. .02

SUBSTITUTION THROUGH AMINO LINKAGE

23632 Propionitrile, 3-hydrazino-, monosulfate.

26057 Propionitrile, 3-(methylamino)-

25451 Propionitrile, 3-(dimethylamino)-

23944 1-Aziridinepropionitrile

13185a Propionitrile, 3-(diethylamino)- .

32938 Propionitrile, 3-(dipropylamino)-

32937 Propionitrile, 3-(dibutylamino)-. . ......

.04

.06

See footnotes at end of table.
HOUSE FLY ATTRACTANTS AND ARRESTANTS 13

TABLE 7.—Arrestant activity of propionitrile derivatives substituted on S position in petri-dish tests—Con.

Code No,

(ENT-) Chemical

SUBSTITUTION THROUGH AMINO LINKAGE—Con.

20446 Propionitrile, 3-(dipentylamino)-

24580 Propionitrile, 3-(octadecyianiino)-

24579 Propionitrile, 3-[(3,5,5-trimethylhexyl)amino]-

16612 Glycine, Ar-(2-cyanoethyl)-

17126 DL-Alanine, Ar-(2-cyanoethyl)-

17125 DL-Aspartic acid, Ar-(2-cyanoethyl)-

20448 Propionitrile, 3-(benzylamino)-

Class

Ratio

T/Std

.03

1 .01

10

1 .04

,02

0

.04

SUBSTITUTION THROUGH KETONE LINKAGE

11714 Propionitrile, 2-benzoyl- .01

1 Two or more tests.

TABLE 8.—Arrestant activity of various chemicals with two or more substitutions on propionitrile molecule

in petri-dish tests

Code No.

(ENT-) Chemical Class

Ratio

T/Std

22936 Lactonitrile, 2-methyl-, carbanilate

17792 Propionitrile, 3-(p-chlorobenzoyl)-2-2-[(3,4-methylenedioxy)phenyl]-

23889 1-Piperidinepropionitrile, alpha-phenyl-^ hydrochloride

23855 Hydratroponitrile, öeto-(benzylamino)-, hydrochloride

23852 Hydratroponitrile, 6e¿a-(dimethylamino)-, hydrochloride

16816 Acrylonitrile, 2,3-diphenyl20206 ^ ^aip/ia-Indoleacetamide, alpha-cyano-2-oxO'

2 0.04

3 .11

2 1 .02

2 12 .05

4 .30

2 .06

3 .25

1 Two or more tests.

2 Seemed to stimulate flies more than ratio would indicate.

However, the compounds in this heterogeneous

group cannot properly be compared with those

chemicals having one substitution. Four chemicals

(ENT-23854, ENT-23889, ENT-23855, ENT23852) are hydrochloride salts bound para to the

phenyl moieties. One of these compounds was in

class 4, whereas the others showed little or no

activity. It will be remembered that three of the

compounds in table 7, which incorporated chlorine

in the para or meta position, were active.

2,3-Diphenylacrylonitrile (ENT-16816) exhibited a small amount of activity but was less

active than 2-phenylacrylonitrile (ENT-7248),

which in turn was less active than acrylonitrile,

whether rated by ratio or class grouping, alphaPhenyl-2-furanacrylonitrile (ENT-17203) differs

from ENT-16816 only in the presence of a furan

ring replacing a phenyl. With this addition all

activity was lost, as was true for 2-furanacrylonitrile (ENT-28307). Thus at least for acrylo^

nitrile and its derivatives, furan rings eliminate all

attractiveness.

CHACON O

ENT-16816

O

CH=CCN

I

O

\n/

ENT-17203

It is difficult to evaluate the overall effect of

substituent groups on a propionitrile molecule. In

general, it seems that the more complex the substituent groups, the less the activity that can be

expected. However, a compound in which the substitution included both a naphthalene group and a
14 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

hydroxyl group, 2-hydroxy-l-naphthalene propionitrile (ENT-16234), was a class 5 arrestant

(ratio 0.83).

CH2CH0CN

ENT-16234

Twenty-two chemicals were available that had

two or more propionitrile moieties bound to larger

molecules. This group of chemicals is listed in

table 9. The relation of structure to activity of the

chemicals containing two propionitrile moieties

is unclear. For example, 3,3'-[[(2-hydroxy-lnaphthyl)methyl]imino]dipropionitrile (ENT22646) differs from 3,3'-[(5-phenylsalicyl)imino]-

dipropionitrile (ENT-22307) only in the type of

CH2CH2CN CH2CH2CN

CH2N

I \

/X/X^OH CH2CH2CN

010

CH2N

/\0H CH2CH2CN

O O

ENT-22646 ENT-22307

cyclic moiety, yet one is class 5 and the other class

1, and 3,3'-(methylimino)dipropionitrile (ENT26303), which has no cyclic moiety, has no

CH2CH2CN

H3CN

CH2CH2CN

ENT-26303

activity. It seems that a hydroxyl group in combination with a naphthalene ring may induce some

activity not entirely attributable to the iminodipropionitrile moiety (ENT-16234 and ENT22646). The imino linkage per se does not appear

to confer activity.

Four chemicals having two propionitrile moieties not bound by an imino-type linkage were

tested and all had class 4 or 5 activity. Included

in this highly active group are A/^,A/'-bis(2«cyanoethyl)-m-toluamide (ENT-26949), which binds

the two propionitrile moieties through an amide

nitrogen, and the ethyl ester of 2,2-bis(2-cyanoethyl) acetoacetic acid (ENT-16032), where they

are bound through an acyl carbon and may be

considered a derivative of pimelonitrile.

0

CH2CH2CN

0

CH3CH2O—C—C—C—CH3

I

CH2CH2CN

ENT-16032

The data concerning the activity of those chemicals with three or four propionitrile moieties are

more clear. 4-Benzoyl-4-(2-cyanoethyl)heptanedinitrile (ENT-16027) exhibits strong arrestancy;

however, any para addition to the phenyl, whether

a methyl, bromine, or chlorine, removes much of

the activity. This is in contrast to the effect of the

para-chlorine additions discussed earlier in connection with other chemicals. The addition of two

chlorine atoms in the meta and para positions

further decreases the arrestancy of this group of

chemicals. Activity in this group is not dependent

on a cyclic moiety, since 4-nitro-4-(2-cyanoethyl)-

heptanedinitrile (ENT-23096) was active. The

two chemicals containing four propionitrile moieties demonstrated only a weak ability to arrest

the flies even though they possess cyclic moieties.

0 CH2CH2CN

/ /

/XC—C—CH2CH2CN

o CH2CH2CN

ENT-16027

CH2CH2CN

/

O2NC—CH2CH2CN

\

CH2CH2CN

ENT-23096

Derivatives of acetonitrile, butyronitrile, and

valeronitrile also demonstrated an ability to arrest

flies; however, not as many derivatives of these

chemicals were available for testing. The results of

tests with active derivatives of acetonitrile are

presented in table 10. The inactive compounds

were as follows:

Code No.

(ENT-) Chemical

23725 Acetonitrile, (4-hydroxy-3-methoxyphenyl)-,

acetate

23875 Acetonitrile, (p-nitrophenyl)-

4979 1-CycIohexene-l-acetonitrile

4980 ^ ^ aipfta-Cyclohexaneacetonitrile, 2-allyl26061 1-Naphthaleneacetonitrile

16855 Aeetonitrile, (phenylsulfonyl)-

23764..... 1-Pyrrolidineaeetonitrile

23766. . . . . Glycinonitrile, Ar-(l,l,3,3-tetramethylbutyi)-,

hydroehloride

4364 Acetonitrile, benzoyl7187a. . . .Acetic acid, cyano-, cyclohexyl ester
HOUSE FLY ATTRACTANTS AND ARRESTANTS 15

Code No,

(ENT-)—Con. Chemical

4983 ^\ a^2^M-Cyclohexaneacetonitrile, alphaphenyi16132 Acetonitrile, 2-phenyl19354 Acetonitrile, benzoylphenyl2754 Glycinonitrile, N-(p-bromophenyl)-2-phenyl20145 Mandelonitrile, 3,4-(methylenedioxy)-

16855 Cyanomethyl phenyl sulfone

Only one single-substituted derivative of

acetonitrile that contained a phenyl group,

(3,4-dimethoxyphenyl)acetonitrile (ENT-23882),

showed relatively strong activity. However, (4-

hydroxy-3-methoxyphenyl)acetonitrile acetate

(ENT-23725) differs only in one acetate group

and exhibits complete lack of stimulation. Although 1-naphthaleneacetonitrile (ENT~26061)

was not active, 2-hydroxy-l-naphthalenepropionitrile (ENT-16234) exhibited marked stimulaH3CO

/XCHaCN

O

0 /XCHsCN

H3CCO O

0CH3

ENT-23882

OCH3

ENT-23725

OP

E^T-26061

tion. Another interesting relationship was noted.

Although cyanomethyl phenyl sulfone (ENT16855) demonstrated no activity, its close relative,

2-cyanoethyl phenyl sulfone (ENT-14259) (table

7), was strongly active. From the evidence presented so far it appears that not only chain length

of the cyanide parent is involved in activity but

also derivatives on the substituent groups.

0 0

\/

/\S—CH2CN

O

0 0

\/

/\SCH2CH2CN

O

ENT-16855 ENT-14259

TABLE 9.—Arrestant activity of various chemicals containing two or more propionitrile moieties

in petri-dish tests

Code No.

(ENT-) Chemical Class

Ratio

T/Std

TWO PROPIONITRILE MOIETIES

26949 m-Toluamide, iV,Ar-bis(2-cyanoethyl)-

16032 Acetoacetic acid, 2,2-bis(2-.cyanoethyl)-, ethyl ester

22646 Propionitrile, 3,3'-[[(2-hydroxy-l-naphthyl)methyl]imino]di17133 DL-Methionine, A/',IV-bis(2-cyanoethyl)-.

15102 Carbamic acid, (2-cyanoethyl)ethyldithio-, zinc salt

16825 Carbamic acid, ethylenebis[(2-cyanoethyl)dithio-, disodium salt.

16878 Carbamic acid, ethylenebis[(2-cyanoethyl)dithio-, zinc salt

17122 DL-Alanine, A/',iV-bis(2-cyanoethyl)-

13041 Malonic acid, bis(2-cyanoethyl)-, diethyl ester

26303 Propionitrile, 3,3'-(methylimino)di24647 Propionitrile, 3,3'-[(3,5-dichlorosalicyl)imino]di22307 Propionitrile, 3,3'-[(5-phenylsalicyl)imino]di17121 DL-Aspartic acid, Ar,N-bis(2-cyanoethyl)- . , 17237 Bisulfide, bis[(2-cyanoethyl)ethylthiocarbamoyl]- , MORE THAN TWO PROPIONITRILE MOIETIES

16027 Heptanedinitrile, 4-benzoyl-4-(2-cyanoethyl)-

16033 Heptanedinitrile, 4-(2-cyanoethyl)-4-23-toluoyl16030 Heptanedinitrile, 4-(p-chlorobenzoyl)-4-(2-cyanoethyl)-

16034 Heptanedinitrile, 4-(í9-bromobenzoyl)-4-(2-cyanoethyl)-

16089 Heptanedinitrile, 4-(2-cyanoethyl)-4-(3,4-dichlorobenzoyl)-

23096 Heptanedinitrile, 4-(2-cyanoethyl)-4-nitro16026 1,1,3,3-Cyclopentanetetrapropionitrile, 2-oxo16028 1,1,3,3-Cyclohexanetetrapropionitrile, 2-oxo5 5.34

5 1.29

5 .79

3 ,15

2 1 .05

2 .03

2 .02

2 2 .02

2 .02

1 0

1 0

1 0

1 0

1 10

5 3.18

4 .52

4 .45

4 .25

4 .18

4 .19

4 .09

3 .07

1 Seemed to stimulate flies more than ratio would indicate.

2 Two or more tests.
16 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

TABLE 10,—Arrestant activity of various substituted acetonitrile derivatives in

petri-dish tests

Code No.

(ENT-) Chemical

Ratio

Class T/Std

SINGLE SUBSTITUTIONS

23882 .Acetonitrile, (3,4-dimethoxyphenyl)- 4

4975 Acetonitrile, phenyl- 2

1T751 Glycinonitrile, iV-methylene- 4

23466 .Acetic acid, cyano-, hydrazide 3

2755 Glycinonitrile, iV-phenyl- 2

2824 — . .Glycinonitrile, iV-(o-methoxyphenyl)- 2

MORE THAN ONE SUBSTITUTION

17436 Acetonitrile, diphenyl- 33

2757..... .Glycinonitrile, N-l-naphthyl-2-phenyl- 3

2753 Glycinonitrile, N,2-diphenyl- 2

17359 Acetonitrile, bis[p-(dimethylamino)phenyl]phenyl~ 3

28712 Acetonitrile, iminodi-. 2

23663 Acetonitrile, (ethylenedinitrilo)tetra- 3

202Q5 3,3-Indolediacetic acid, alpha.alpha' dicyano-2-oxo-, diethyl ester. 2

10.25

1 .01

.45

.11

.01

2 .05

(13)

.18

.03

.31

.01

2 .56

2 .14

1 Two or more tests.

- Seemed to stimulate flies more than ratio would indicate.

^ Induced greatly different activities in two tests: 0:27; 38:292.

Certain substitutions linked through nitrogen

seemed to possess activity, although not strongly.

The most potent arrestant among this group was

iV-methyleneglycinonitrile (ENT-20150). This

chemical resembles, and was as effective as, acrylonitrile (ENT-54). Other members of this group

demonstrated a consistent but low activity. Two

other chemicals were placed arbitrarily in this

series, benzoylacetonitrile (ENT-4364) and cyclohexyl cyanoacetate (ENT-7187a). Neither of

these exhibited any activity.

H2C = NCH2CN

ENT-20150

Several double- and triple-substituted derivatives of acetonitrile were relatively good arrestants, but none were good enough to be rated in

class 4 or 5. Iminodiacetonitrile (ENT-60248) exhibited very slight activity, even though two

acetonitrile moieties were present.

CH2CN

HN

CH2CN

ENT-60248

Isobutyronitrile (ENT-28525) was a class 4

arrestant of great variability with a ratio of 0.04.

Only one of the single-substituted derivatives of

butyronitrile that was tested, 2-cyanobutyramide

(ENT-20170), showed any arrestancy (class 2,

ratio 0.08). The inactive compounds were as

follows:

Code No.

(ENT-) Chemical

19356 Butjo-onitrile, 4-(diethylamino)-

16701 Butyronitrile, 4-(2,4-dichlorophenoxy)-

7451 Butyric acid, 2-cyano-, cyclohexyl ester

7464. . . . .Butyric acid, 2-cyano-, 2-methylcyclohexyl ester

7467 Butyric acid, 2-cyano-, 3-methylcyclohexyl ester

7468 Butyric acid, 2-cyano-, 4-methylcyclohexyl ester

26028. Butyronitrile, 4-(octylthio)-

26029 Butyronitrile, 4-(octylsulfinyl)-

23765. . . . . Propionitrile, 2,2'-(ethylenediimino)bis[2-

methylA few of these derivatives were similar to propionitrile derivatives, which showed no activity

(cf. 4-(diethylamino)butyronitrile (ENT-19356)

with 3-(diethylamino)propionitrile (ENT-13185)).

Although tests of these single-substituted derivatives failed to provide much information, tests of

those with double-, triple-, and quadruple-substituted molecules provided some meaningful in-
HOUSE FLY ATTEACTANTS AND ARRESTANTS 17

formation. These are presented in table 11. Isopropylcyanoaeetamide ( = 2-cyano-3-methylbutyramide) (ENT-20149) was a strong arrestant.

Some activity might have been expected from 4-

(p-chlorobenzoyl) - 2 - cyano - 3 - phenylbutyrate

(ENT-16067) because of the chlorine in the para

position. However, the close proximity of the

carboxyl group to the cyanide radical probably

eliminated this expected activity.

0

H3C V

NH2

CH2CCN

' H

H3C

ENT-20149

The most interesting information and perhaps

the most significant concerns the different isomers

of butyronitrile derivatives separated by differences in melting points. Two isomers of 4-benzoyl2,3-diphenyl butyronitrile (ENT-17780) were

tested, one of which melted at 109° C. (ENT17779) and the other at 117° (ENT-17780). The

isomer melting at the lower temperature was a

class 5 arrestant, whereas the isomer melting at

117° was in class 3.

/\

O

0

y\c—CH2CHCHCN

P A ^ (ol

ENT-17779

Another rather close relative of this chemical

is 4-benzoyl-2-bromo-2-(p-nitrophenyl)-3-phenylbutyronitrile (ENT-17794). Two isomers were

tested, one with a melting point at 151° C. (ENTO

o X/Br

/\C—CH2CHCCN

o) À

o

N02

ENT-17794

17793) and the other at 191° (ENT-17794). Again

the isomer melting at the lower temperature was

included in class 5, whereas the isomer melting at

the higher temperature was a class 1 arrestant.

A reversal seemed to occur when two isomers

of 4-benzoyl-2-(p-nitrophenyl)-3-phenyl butyronitrile (ENT-17777 and ENT-17778) were tested.

The chemical did not prove to be a good arrestant,

although lacking only the bromine of ENT17793. However, the isomer with the 170° C. melting point (ENT-17777) was a weak class 3 arrestant, whereas the isomer with the 144° melting

point (ENT-17778) was in class 1.

/ r

/\C—CH2CHCHCN

PJ APi

NO2

ENT-17777

These few data do not prove that arrestancy is

dependent on low melting or boiling points. However, these three chemicals do definitely illustrate

that stereochemical configuration is important to

activity as arrestants.

A small number of valeronitrile derivatives were

available for screening. Since the parent chemical

was ineffective (table 6), little activity was expected from this group of chemicals. However, the

three chemicals having an amide group were effective—2-cyanovaleramide (ENT-20148, class 5,

ratio 1.18), 2-cyano-4-methyl-2-propylvaleramide

(ENT-20209, class 3, ratio 0.07), and 2-cyano-2-

isopropylvaleramide (ENT-20210, class 2, ratio

0.02). The inactive compounds were as follows:

Code No.

(ENT-) Chemical

20151 Valeronitrile, 5-chloro11712 Valeronitrile, 2-benzoyl16512 Valeronitrile, 5,5'-oxydi23095 Valeronitrile, 4-methyl-4-nitro22305 Heptanedioic acid, 4,4-dicyano-, diallyl ester

7351 Valeronitrile, 4-hydroxy-2-phenyl13167 Glutaric acid, 2-cyano-3-methyl-, diethyl ester

13172. . . . .Glutaric acid, 2-cyano-3-metliyl-, allyl ethyl

ester

17776 Hydrocinnamic acid, 2-cyano-3-phenacyl-,

methyl ester

4981 1-Cyclohexene-1-acetonitrile, 2-allyl-
18 AGRICULTURE HANDBOOK 403, U.S. DEPT. OP AGRICULTURE

TABLE 11.—Arrestant activity of multiple-substituted derivatives of butyronitrile

in petri'dish tests

Code No.

(ENT-) Chemical

Ratio

Class T/Std

TWO SUBSTITUTIONS

20149 Butyramide, 2-cyano-3-methyl- 5

9216 Butyronitrile, 2-]iydroxy-2-methyl-, acetate 1

THREE SUBSTITUTIONS

(p-chlorobenzoyl)-2-cyano-3-phenyl- 1

;-(j)-methoxybenzoyl)-2,3-diphenyl- 1

•benzoyl-2,3-diphenyl- (isomer m.p. 109° C.) • 5

:-benzoyl-2,3-diphenyl- (isomer m.p. 117°C.).... 3

4-benzoyl-2-(p-nitrophenyl)-3-phenyl- (isomer

4-benzoyl-2-(2?-nitrophenyl)-3-phenyl- (isomer

16067 Butyric acid, 4-

17663 Butyronitrile, 4

17779 Butyronitrile, 4

17780 .Butyronitrile, 4

17778 .Butyronitrile,

m.p. 144° C.)

17777 .Butyronitrik

m.p. 170° C.) 3

5611 .Succinic acid, 2-cyano-3-methyl-, diethyl ester 1

5645 Succinic acid, 2-cyano-2,3-dimethyl-, diethyl ester 3

FOUR SUBSTITUTIONS

17672 Butyronitrile, 4-benzoyl-2-bromo-2,3-diphenyl-. 4

17793 Butyronitrile, 4-benzoyl-2-bromo-2-(p-nitrophenyl)-3-phenyl-

(isomer m.p. 151° C.) 5

17794 Butyronitrile, 4-benzoyl-2-bromo-2-(p-nitrophenyl)-3-phenyl-

(isomer m.p. 191° C.) 1

0.70

0

0

0

.67

.05

.05

)

.10

.29

.60

Two or more tests.

H3C CH2CH2CH3

H3CCH2CH2CHCN CHCH2C—CN

I / I:

C=0 H3C c=o

I I

NH2 NH2

ENT-20148 ENT-20209

HgC CH3

\ /

CH

I

H3CCH2CH2CCN

I

C=-0

I

NH2

ENT-20210

The next large group of chemicals that were

screened were those having one or more cyano

groups bound directly to a cyclic moiety. Results

with the active compounds are given in table 12.

The inactive compounds were as follows:

Code No,

(ENT-) Chemical

24184 Benzonitrile

492. . ... Benzonitrile, p-chloro33363 Benzonitrile, 2,3-dichloro17128 o-Tolunitrile

17129 m-Tolunitrile

14826 :p-Tolunitrile

488a.... Benzonitrile, p-bromo11801 p-Anisonitrile

11298 Benzonitrile, 2,4,6-triisopropyl25034 Isophthalonitrile

234 2-Naphthonitrile

22124 9-Phenanthrenecarbonitrile

23919. . . .. 9-Fluorenecarbonitrile, 9-amino5928..... 5-Norbornene-2-carbonitrile

16843. . . . .5-Norbornene-2,3-dicarbonitrile

26364 7-Oxabicyclo[4.1.0]heptane-3-carbomtrile

5832 .....Butyric acid, 2-cyano-2-phenyl17235 Nicotinonitrile, l,2-dihydro-4,6-dimethyl-2-oxo1547 Nicotinonitrile, l,2-dihydro-l,4,6-trimethyl2-0X0-

8635 3-Cyclohexene-l-carbonitrile

3957 Cyclohexanecarbonitrile, l-hydroxy-x-methyl

1651. . .. . 3-Cyclohexene-l-carbonitrile, 5,5-dimethyl-2-

(1-methylpropenyl)-

21026. . ., .Butyramide, iV-(l-cyanocyclohexyl)-iV-methyl-
HOUSE FLY ATTRACTANTS AND ARRESTANTS 19

TABLE 12.—Arrestant activity of chemicals having one or more cyanide radicals bound directly to cyclic

moieties in petri-dish tests

Code No.

(ENT-) Chemical Class

Ratio

T/Std

33327 Benzonitrile, 2,4,5-trichloro33325 Benzonitrile, 2,4,6-trichloro478 Benzonitrile, p-nitro49 Phthalonitrile

16842 5-Norbornene-2,3-dicarbonitrile, 2-methyl16848 7-Oxabicyclo[2.2.1]hept-5-ene, 2,3-dicarbonitrile, l-(hydroxymethyl)-, acetate. . . , 16983 2,3-Norbornanedicarbonitrile, 5,6-dichloro-, alpha-isomer

25962 2-Norbornanecarbonitrile, exo-5-chloro-6-oxo-, 0-(methylcarbamoyl)oxime, endo-.

18124 Cyclohexanecarbonitrile, 1-heptyl21025 Propionamide, iV-(l-cyanocyclohexyl)-iV-methyl16844 4-Cyclohexene-l,2-dicarbonitrile, 3,3-dimethyl-6-(l-methylpropenyl)-

3

2

3

4

2

2

2

4

4

3

2

^0.07

1 .02

.02

.13

1 .10

i .08

12 .05

.17

.23

.27

1 .02

1 Two or more tests.

2 Seemed to stimulate flies more than ratio would indicate.

The least complex member of this group was

benzonitrile (ENT-24184), which evoked no response from the flies. Since it was earlier

hypothesized that a chlorine in the para position

seemed to enhance activity, p-chlorobenzonitrile

(ENT-492) and 2,3-dichlorobenzonitrile (ENT33363) were screened and found inactive. However, two trichlorobenzonitrile derivatives evoked

a small positive response. Methyl groups in any

position did not confer any activity, but a paranitro addition evoked a class 3 response.

/XCN

O

ENT-24184

Phthalonitrile (ENT-49) was active, but isophthalonitrile (ENT-25034) was totally inactive.

5-Norbornene-2,3-dicarbonitrile (ENT-16843)

was inactive, but the addition of a methyl

group in 2-methyl-5-norbornene-2,3-dicarbonitrile

(ENT-16842) restored some of the activity. Although chlorine substitution conferred no activity

to benzonitrile, two chlorine additions to the

norbornene ring to produce 5,6-dichloro-2,3-norbornanedicarbonitrile (ENT-16983) restored some

activity to the inactive molecule. 1-Heptylcyclohexanecarbonitrile (ENT-18124) evoked a class 4

response, even with the attachment of a seven

carbon chain as did eíco-5-chloro-6-oxo-eticío-2-

norbornanecarbonitrile 0-(methylcarbamoyl)oxime (ENT-25962).

O

ENT-49

CN

O

\/CN

ENT-25034

CHs

CN

CH2

CN

ENT-16842

CN

CH2

CN

ENT-16843

Cl CN

CH2

Cl CN

ENT-16983

Table 13 lists some effective long carbon-chain

nitriles. Generally these chemicals demonstrated

little to no activity. The inactive compounds were

as follows:

Code No.

(ENT-) Chemical

4913 3-Pentenoic acid, 2-cyano-3-ethyl-2-propyl-,

ethyl ester

4912 3-Pentenoic acid, 2-cyano-2,3-diethyl-, ethyl

ester

4965 3-Pentenoic acid, 2-butyl-2-cyano-3-methyl-,

ethyl ester

4916 2-Hexenoic acid, 2-cyano-3-methyl-, ethyl ester

4915 2-Hexenoic acid, 2-cyano-3,5-dimethyl-, methyl

ester

4914 2-Hexenoic acid, 2-cyano-3-isobutyl-5-methyl-,

ethyl ester

4963 3-Heptenoic acid, 2-cyano-2,3-dimethyl-, ethyl

ester

28306 2-Octenenitrile, 5,7,7-trimethyl4918 2-Octenoic acid, 2-cyano-3-methyl-, methyl ester

4920 2-Octenoic acid, 2-cyano-3-pentyl-, ethyl ester

4911 3-Octenoic acid, 2-cyano-2,3-dimethyl-, ethyl

ester

28305 2-Nonenenitrile

22306 Octadecanenitrile, 9,10-epoxy22318 Octadecanenitrile, ??-tolyl-
20 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

TABLE 13.—Arrestant activity of various long-chain nitriles in petri-dish tests

Code No.

(ENT-) Chemical

Ratio

Class T/Std

7106...... Hexanenitrile, 2,4-dihydroxy-3,3,5-trimethyl20211 Hexanamide, 2-cyano-5-methyl-2-propyl- . . . .

4957 3-Pentenoic acid, 2-cyano-3-methyl-2-propyl-, ethyl ester.

23623 Heptanenitrile, 2-hydroxy-4,6,6-trimethyl-, carbanilate...

28304 2-Decenenitrile

22317 Octadecanenitrile, 10-phenyl- .

22319 Octadecanenitrile, tolylene, di3 iQ.ll

3 1 .19

2 .01

2 1 .01

3 .08

2 1 .08

2 1 .05

1 Two or more tests.

Only two phosphorus esters with various nitriles

demonstrated even slight activity. These two were

0,0-dimethyl 0-phosphorothioate, 0-ester with

4-hydroxy-m-anisonitriIe (ENT-27230, class 2,

ratio 0.08) and 0,0-dimethylphosphorothioate, 0-

ester with 5-chlorosalicylonitrile (ENT-27235,

class 2, ratio 0.02). The inactive compounds

were as follows:

Code No.

{ENT~) Chemical

16155 Phosphonic acid, (2-eyanoethyl)-, diethyl ester

16156 Phosphonic acid, (2-cyanoethyl)-, dibutyl ester

27175 Phosphonic acid, (4-cyanobutyl)-, diethyl ester

27174. . . . .Phosphonic acid, (4-cyanobutyl)-, ethylp-nitrophenyl ester

25869 Phosphonothioic acid, ethyl-, 0-2-chloroethyl

ester, 0-ester with p-hydroxybenzonitrile

27033 Phosphonothioic acid, methyl-, 0-ethyl ester,

0-ester with p-hydroxybenzonitrile

25832. , . . .Phosphonothioic acid, phenyl-, 0-ethyl ester,

0-ester with p-hydroxybenzonitrile

27227 Phosphonothioic acid, (chloromethyl)-, 0-ethyl

ester, 0-ester with 4-hydroxy-m-anisonitrile

27028 Phosphonothioic acid, (chloromethyl)-, O-ethyl

ester, 0-ester with p-hydroxybenzonitrile

The chemicals listed in table 14 are derivatives

of various dicyano compounds given in table 6.

Only one of the malononitrile derivatives, [4-[bis-

(2 - chloroethyl)amino] - 2 - methylbenzylidene]-

malononitrile (ENT-50199), exhibited unquestionable arrestancy.

Several related derivatives of succinonitrile were

available that provide some insight into the effect

of slight molecular changes on arrestancy. Although phenyl succinonitrile (ENT-16982) was a

class 2 arrestant, 2,3-diphenyl succinonitrile

(ENT-25341) was a class 5 arrestant. The addition

of a methylenedioxy (ENT-25343) or methoxy

(ENT-25346) group to one of the phenyl rings of

ENT-25341 diminished the activity to class 4.

The addition of chlorine in the para position

caused a further reduction in activity to class 3

(ENT-25342 and ENT-25344). Two para chlorine

atoms (ENT-25340) eliminated most of the

activity. A para isopropyl group (ENT-25345)

was about as active as a chlorine substitution.

NCCHCH2CN

O

ENT-16982

CN—CH—CH—CN

O O

0

CH2

ENT-25343

CN—CH—CH—CN

O o

Cl OCH3

ENT-25342

CN—CH—CH—CN

/ \

^ /\ 01 (o

01 Cl

CN—CH—CH—CN

/ \

O o

ENT-25341

CN—CH—CH—CN

O O

0CH3

ENT-25346

CN—CH—CH—CN

O o

Cl

ENT-25344

CN—CH—CH—CN

/ \

o O

ENT-25340

CH

/ \

II3C CII3

ENT-25345
HOUSE FLY ATTRACTANTS AND ARRESTANTS 21

TABLE 14.—Arrestant activity of i chemicals with 2 cyano radicals in petri-dish tests

Code No.

(ENT-) Chemical Class

Ratio

T/Std

16838..... .Malononitrile, 1-cyclopropylethylidene19689 Malononitrile, benzylidene16815 Malononitrile, (aZp/ia-methylbenzylidene)-

16819 Malononitrile, (p-alpha-dimethylhenzylidene)-

16820..... .Malononitrile, (m-ethyl-aZpÄa-methylbenzylidene)-

16821 Malononitrile, (aíí5^G-methyl-í?-phenylbenzylidene)-

50199 Malononitrile, [4-[bis(2-chloroethyl)amino]-2-methylbenzylidene]-

24628 Tartrononitrile, methyl-, acetate

23333 Succinonitrile, tetramethyl16982...... Succinonitrile, phenyl25341 Succinonitrile, 2,3-diphenyl25344 Succinonitrile, 2-(??-chlorophenyl)-3-phenyl25340 Succinonitrile, 2,3-bis(p-chlorophenyl)-

25342 Succinonitrile, 2-(î>-chlorophenyl)-3-(p-methoxyphenyl)-

25346 . .Succinonitrile, 2-(î?-methoxyphenyl)-3-phenyl25345 Succinonitrile, 2-p-cumenyl-3-phenyl25343 Succinonitrile, 2-[3,4-(methylenedioxy)phenyl]-3-phenyl14265 4-Morpholinesuccinomtrile

13063 Heptanedinitrile, 4-acetyl-4-methyl16090 Heptanedinitrile, 4-benzoyl-4-methyl1 0

1 0

2 .09

1 10

2 .01

2 1 .06

3 .12

1 0

1 0

2 .02

5 1 .54

3 1 .06

2 1 .03

3 1 .15

4 1 .19

3 1 .09

4 1 .59

3 .05

4 1 .28

4 .89

1 Two or more tests.

The best arrestancy for this entire group was

exhibited by 4-benzoyl-4-methyl-heptanedinitrile

(ENT-16090) and 4-acetyl-4-methyl-heptanedinitrile (ENT~13063). This is an interesting

phenomenon in view of the fact that heptanedinitrile (ENT-28713) by itself was inactive.

However, by comparing these two chemicals with

O

o

c—cCH2CH2CN

-CH3

0

H3CC—cCH2CH2CN

-CH3

CH2CH2CN

ENT-16090

CH2CH2CN

ENT-13063

some of those in table 9, the similarity apparent in

the heptanedinitrile molecule per se becomes less

important than the structural similarity to the

dipropionitrile derivatives. This similarity then

most probably explains the activity of these two

heptanedinitrile derivatives although the parent

chemical was inactive.

Nine miscellaneous compounds were also tested.

Cyanoguanidine (ENT-14632) was a class 3 compound (ratio 0.34). The following compounds were

either ineffective (class 1) or slightly effective

(class 2, ratios 0.02-0.03):

Code No.

{ENT-) Chemical

22316.....Cyanamide, dibenzyl22314..... Cyanamide, bis(2-methylallyl)-

17789 Cyanamide, diallyl8193 s-Triazine, 2,4,6-trimethoxy8192 s-Triazine, 2,4,6-tributoxy24917, . . . .1,3,5,2,4,6-Triazatriphosphorine, 2,2,4,4,6,6-

hexachloride

16307. . . . .Urea, l-acetyl-3-cyano22927. . . . .Acetamide, iV-(cyanoamidino)-

The data and chemical information previously

discussed are indicative of only general trends. No

absolute rule on the structural relationship to

activity could be obtained with the limited number of similar chemicals available for testing.

However, an abbreviated form of ''nearest neighbor'' or active-group-position analysis was obtained for most of the chemicals tested excluding

the thiocyanate compounds. One such analysis

was made of compounds containing a cyano radical

in various positions with respect to a carboxyl or

ester group. The number of compounds in each

class at each position was as follows:
22 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

Position

lass alpha beta gamma delta epsilon

1. . . .. 60 2 7 6 2

2... 7 0 2 3 0

3. . . 3 1 0 1 0

Although none of the relationships were productive of compounds with a high activity, the percentage of slightly active compounds was a little

higher at the greater distances between the two

radicals.

Fifty of these compounds not included in previous tables are listed below. All were ineffective

or only slightly effective, with ratios of 0.01 to

0.05.

Code No.

(ENT-) Chemical

5599..... Acetic acid, cyano-, methyl ester

19027. . . . .Acetic acid, cyano-, ethyl ester

32563 .....Acetic acid, cyano-, propyl ester

7427 Acetic acid, cyano-, hexyl ester

7380 Acetic acid, cyano-, 2-ethylhexyl ester

7221 Acetic acid, cyano-, 2-ethoxyethyl ester

13014. . . . Acetic acid, cyano-, 2-(2-butoxyethoxy) ethyl

ester

7370b .... Acetic acid, cyano-, 3-methylcyclohexyl ester

7371 Acetic acid, cyano-, 4-methyleyclohexyl ester

19051 Acetic acid, cyano (2,4,6-trimethylphenyl)-

23778 Acrylic acid, 2-cyano-3-ethoxy-, ethyl ester

7457 Propionic acid, 2-cyano-, cyclohexyl ester

4908 2-Pentenoic acid, 2-cyano-3-methyl-, ethyl ester

4909 2-Pentenoic acid, 2-cyano-3-ethyl-, ethyl ester

11617 Valeric acid, 2-cyano-, ethyl ester ^

11064 Butyric acid, 2-cyano-4-methyl-, ethyl ester

4910 Hexanoic acid, 2-cyano-, ethyl ester

7489 Hexanoic acid, 2-cyano-3,3-dimethyl-4-oxo-,

ethyl ester ^

13044 Hexanoic acid, 2-cyano-3,3-dimethyl-4-oxo-,

allyl ester

13010 Hexanoic acid, 2-cyano-3,3-dimethyl-4-oxo-,

2-methoxyethyl ester

4907 .....Hexanoic acid, 2-cyano-2-isopropyl-, ethyl ester

4931 Sorbic acid, 2-cyano-3,5-dimethyl-, methyl ester

4960 3-Hexenoic acid, 2-cyano-2,3,5-trimethyl-,

methyl ester

4964 3-Hexenoic acid, 2-cyano-2-ethyl-3,5-dimethyl-,

methyl ester ^

4962 3-Hexenoic acid, 2-cyano-3-methyl-2-propyl-,

ethyl ester

4959 3-Hexenoic acid, 2-cyano-2-ethyl-3-propyl-,

ethyl ester

7224 Octanoic acid, 2-cyano-4-ethyl-, methyl ester

4995. . . . Octanoic acid, 2-cyano-3-methyl-, ethyl ester

Code No.

(ENT-) Chemical

4997 Nonanoic acid, 2-cyano-, ethyl ester

4996 Nonanoic acid, 2-cyano-3-methyl-, ethyl ester

4999 Cyclohexaneacetic acid, alpha-cysLUo-, ethyl ester

7622 Cyclohexaneacetic acid, aip?ia-cyano-4-

methoxy-, ethyl ester

7020 ^ S aZp/ia-Cyclohexaneacetic acid, alpha-cyano-f

ethyl ester

7170 Cyclohexaneacetic acid, alpha-cyano-, propyl

ester

7184 Cyclohexaneacetic acid, alpha-cyano-^ allyl ester

7175 Cyclohexaneacetic acid, alpha-cyano-, isopropyl

ester

7226 Cyclohexaneacetic acid, alpha-cyano-, 2-

ethoxyethyl ester

7185..... Cyclohexaneacetic acid, alpha-cyano-^

cyclohexyl ester

7240 Cyclohexaneacetic acid, alpha-cyano-^ benzyl

ester

24083 .... . Cinnamie acid, p-chlovo-alpha-cyano- ^

10525 Cinnamie acid, alpha-cyano-, ethyl ester

4998..... Hydrocinnamic acid, alpha-cyano-, ethyl ester

4925. . . . .Cinnamie acid, alpha-cyano-heta-methyl-, ethyl

ester

4928 Cinnamie acid, alpha-cyano-beta-eth.yl-f ethyl

ester

4934 Cinnamie acid, a^p/ia-cyano-6e¿a-propyl-, ethyl

ester

4926 Cinnamie acid, alpha-cyano-beta-pentyl-, ethyl

ester

20317 Cinnamie acid, aip/^a-cyano-o-methoxy-, ethyl

ester ^

8518 Cinnamie acid, alpha-cyano-p-methoxy-f ethyl

ester ^

20318 Cinnamie acid, a^j?/ia-cyano-3,4-(methylenedioxy)-, ethyl ester ^

15927 Pyruvic acid, 3-cyano-3-phenyl-, ethyl ester i

1 Class 2 or 3.

An analysis was also made of compounds containing a cyano radical separated by various numbers of carbon atoms from a carbon-carbon double

bond. The number of compounds in each class at

each distance was as follows:

Distance

lass 0 1 2 S

1 22 5 5 1

2 5 0 1 0

3 2 0 0 0

4 1 1 0 0

The group included only four compounds in classes
HOUSE FLY ATTRACTANTS AND ARRESTANTS 23

3 and 4, and these were all within one carbon of

the double bond.

The following results were obtained with compounds in which a nitrogen atom was located in

various positions relative to a cyano radical :

Position

Class ^ 0 alpha beta gamma

1 4 6 14 4

2 2 4 11 0

3 1 4 10 0

4 0 0 2 1

5 0 0 4 0

1 Cyanide radical bonded directly to nitrogen.

Thirty-nine, or 58 percent, of these 67 chemicals

exhibited some degree of activity (class 2 or

better) ; 27 of the 41 with the nitrogen in the beta

position, or 66 percent, were active, and these

included six of the seven most effective compounds

(classes 4 and 5). These proportions indicate that

a nitrogen in the beta position is at least conducive

to activity.

Of five chemicals tested with a sulfur located on

the beta carbon from the cyanide, three were

active; two of these had activity above class 3.

Evaluation of the effect of acyl and amide

groups was also made, with the following results:

Position

Class alpha heWT gamma

1 4 J. 2

2 4 0 2

3 4 4 3

4 0 0 7

5 2 0 5

As seen from these data, almost all compounds

with an acyl or amide group in the gamma position were active. Of course, the fact that most of

these are dipropionitrile derivatives must not be

overlooked. Even those chemicals with alpha and

beta acyl or amide groups showed consistent if

slightly lower activity on the average. It seems

safe to say, then, that chemicals that possess

gamma acyl or amide links are the most promising.

The importance of this linkage is further supported by the results with 3,3'-[(3,5-dichlorosalicyl)imino]dipropionitrile (ENT-24647), which

has two propionitrile groups, but no acyl or amide

linkage and no activity. This is also true of ENTCl

Cl

CH2CH2CN

o CH2N

OH CH2CH2CN

ENT-24647

22307, ENT-17121, and a few others. Only

moderate activity was obtained from two tetraHO2CCH2 CH2CH2CN

N

HO2CCH2 CH2CH2CN

ENT-17121

propionitrile derivatives, ENT-16026 and ENT16028. These also contain a saturated ring, which

was usually not associated with high activity.

NCCH2CH2 CH2CH2CN

X.0

~CH2CH2CN

CH2CH2CN

ENT-16026

NCCH,CH2 CH2CH,CN

A-

\/~CH2CH2CN 1

CH2CH2CN

ENT-] 6028

Except among the straight-chain hydrocarbons,

only three chemicals with no acyl or amide linkage

produced a high degree of arrestancy (class 5).

These three chemicals are ENT-16966, ENT16234, and ENT-22646, which are slightly different from similar chemicals showing activity.

Two are naphthalene derivatives containing

hydroxyl groups in different locations. The third

is a para-chloro substituted phenyl derivative

with a thio linkage. None of the other class 5

chemicals are closely related to any of these three

chemicals, and it is possible that the mode of

stimulation by these chemicals is somehow

different from the rest.
24 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

Only 39 chemicals were available in which the

cyano radical was attached directly to a cyclic

structure, and only 11 had activity ranging from

class 2 to class 4 (28.2 percent). This percentage

is lower than that calculated for all chemicals, and

it is thus not unreasonable to consider that these

types of cyanide derivatives will not be as

stimulatory to the house fly as other types.

An analysis was also made of a group of compounds in which a cyano radical was situated at

various distances from a cyclic moiety, i.e.,

separated by various numbers of carbon, nitrogen,

or (in a few compounds) oxygen atoms. The number of compounds in each class at each distance

was as follows:

Distance {carbon atoms)

Class 1 2 3 U

1 33 26 2 9

2. . 11 11 1 1

3 Ö 9 1 3

4 4 3 1 6

5. . 2 3 0 5

Of 56 chemicals having a cyano radical

separated from the cyclic moiety by one atom, 23

were active (41 percent) with all classes represented, almost identical with the percentages

among all compounds tested. All the 13 cyclohexyl

compounds were in class 1. If only the phenyl

compounds are considered, the proportion of

active compounds was 56 percent, substantially

higher than the general average.

The next group to be considered was that where

the cyanide is located two atoms from the cyclic

moiety. In this group, 50 percent of the chemicals

were active with all groups represented, again

higher than the general average, but still slightly

disappointing since this group includes most of the

simple propionitrile derivatives.

The largest proportion of active compounds was

found among those with cyanides located four

atoms from a phenyl ring, 63 percent of which

were in class 2 or higher and 46 percent of which

were in classes 4 and 5. Of course, many compounds in this group had acyl and amide linkages

as discussed previously. Unfortunately few chemicals were available to adequately assess the

importance of the phenyl group compared with the

amides.

CONCLUSIONS

The importance of the foregoing results lies not

only in the demonstration of arrestant activity in

certain compounds but in the indication that certain molecular configurations possessing this

action can possibhy be combined with moieties

having other properties without destroying either

arrestant or other properties. The limited research

with this generic group of chemicals has already

indicated that different members may elicit one

or more of the following behavioral responses:

(1) Attraction, (2) arrestancy, (3) stimulation of

feeding, or (4) repellency. These chemicals should

be more thoroughly studied, since it is conceivable

that any member of the group might affect

different species of insects in different ways.

Acetonitrile, propionitrile, butyronitrile, and

isobutyronitrile are relatively potent arrestants,

and certain derivatives of these basic groups still

retain this activity. It has been shown that the

acyl or amide group, in combination with

propionitrile (II), can be used with some chemical

0

II /

R—C—(N) (N may also = C)

II

groups to produce arrestancy in house flies. If

a chemosterilant moiety could be combined in the

same molecule without impairment of either the

arresting or the sterilizing action, the single compound would be more effective in practical application than the chemosterilant alone, since the insects would remain in contact with it for longer

periods and ingest more of the compound if it is a

feeding stimulant as well as an arrestant.

For example, a combination with hempa (III)

or its relatives such as tepa, metepa, or thiotepa

might prove active in both respects. With apholate

several combinations might be possible, e.g., IV,

However, the activity of the nitriles could be

impaired by proximity to phosphorus. The III
HOUSE FLY ATTRAGTANTS AND ARRESTANTS 25

and IV structures have been used here only

to illustrate a point. More and better nitrile

derivatives may be found.

H2C 0 CH2CH2CN

//

N C—N

CH2CH2CN

N

CH2CH2CN

H3C C=0

\ I

N—P=0

/ I

H3C N

/ \

H3C CH3

III

H2C

H2C

H2C

N

N N

CH2CH2CN

CH2

N

P CH2

CH2

N

N N'

H2C CH2 CH2

IV

SUMMARY

Tests were made in an olfactometer to determine the attractiveness of ammonia and various

nitriles to house flies (Musca domestica L.). Petridish tests were made to determine the arrestancy

of these and related compounds. The olfactometer

tests with ammonia showed that the attraction

conformed to the Weber-Fechner relationship

over four log units of concentration. Olfactory

tests with 52 nitriles were not indicative of

attraction.

A total of 53 toluamide and benzamide derivatives and 390 cyanides, thiocyanates, or isothiocyanates were screened as arrestants on white sand

in competition with granulated sugar. The compounds were placed in classes 1 to 5 according to

the number of flies arrested by the test chemical,

irrespective of the number on the sugar.

Ar,iV-Bis(2-cyanoethyl)-m-toluamide (ENT26949) exerted a strong degree of arrestancy. However, the overall results with the 53 toluamide and

benzamide derivatives indicated these structures

were not responsible for the degree of arrestancy

of this chemical.

Three inorganic cyanides were tested. Two of

them were slightly active as arrestants, indicating

that a cyano radical was responsible for at least

some of the arrestant activity.

Fifty-four thiocyanates were tested to determine the activity of this radical, since the literature suggested that some effect might be expected.

No thiocyanate was found that could be placed in

class 4 or 5. This was also true of eight isothiocyanate compounds.

The activity of iV,Ar-bis(2-cyanoethyl)-m-toluamide (ENT-26949) and tests with the toluamide

and benzamide derivatives suggested that the

propionitrile moiety (ENT-8777) might have been

responsible for the arrestant activity of this chemical. A series of straight-chain aliphatic nitriles was

tested, and three of these—acetonitrile (ENT327), propionitrile (ENT-8777), and butyronitrile

(ENT-8778)—were shown to be strong arrestants,

with propionitrile the best. Activity decreased

abruptly after chain lengths of four carbon atoms.

Seven straight-chain aliphatic compounds having two cyanide radicals demonstrated poor

activity. Acrylonitrile (ENT-54), a single

unsaturated 3-carbon cyanide, showed a definite

but weaker activity than propionitrile.

Generally all substitutions on the 2 position of

propionitrile were less effective than the parent

chemical. Only one chemical with an alpha substitution on a propionitrile molecule had even a

low order of activity. Thirteen chemicals considered to be multiple substitutions on a basic

propionitrile molecule were tested, and four of

these were active.

Fourteen chemicals were tested that had two

propionitrile moieties attached to a single molecule. Generally more of these were active than

any other group, especially those linked through

an amide or acyl group. Only eight chemicals were

tested that had more than two propionitrile moieties on a single molecule. All of these were active.

The best were those with acyl linkages.

Only six single-substituted acetonitrile derivatives were active out of 16 tested, and only three

were class 3 or higher. Only 13 other acetonitrile

derivatives were available for testing, and none

showed activity greater than class 3.
26 AGRICULTURE HANDBOOK 403, U.S. DEPT. OF AGRICULTURE

Eleven single-substituted derivatives of butyronitrile were available for testing. One of these,

isobutyronitrile, possesses definite activity (class

4), Thirteen multiple-substituted butyronitrile

derivatives were tested. Seven of these exerted

strong arrestant activity.

Thirteen derivatives of valeronitrile were tested,

three of which werv.- active. These were the only

ones with amide groups near the cyanide radical.

Thirty-four chemicals were tested that had one

or more cyano radicals bound directly to a cyclic

moiety. These were generally inactive, except

three, which were class 4 arrestants. None

of these three possessed other similar structural

similarities.

Tests were made with 23 nitriles with chain

lengths greater than five carbon atoms, many of

which had the cyanide radical in the alpha positionrelative to an ester linkage. None were better

than class 3 arrestants. Tests with 50 shorter

chain chemicals where the cyano radical was in the

alpha position relative to an ester linkage did not

indicate activity greater than class 2 or 3.

Tests with 11 phosphorus esters with various

nitriles produced only two chemicals in class 2.

Tests with 18 derivatives of malononitrile and

succinonitrile indicated no activity above class 3

except ENT-25341, ENT-25343, and ENT25346. Two pimelonitrile derivatives were class 4

arrestants. Both of these may be considered as

dipropionitriles joined near an acyl group.

It was concluded that three aliphatic nitriles

were highly active as house fly arrestants and that

two or more propionitrile molecules could be

bound to other molecules and at least retain

activity as long as the bond was through an acyl or

amide linkage. Certain combinations with cyano

radicals seemed directly responsible for lack of

activity. The predominant one of these was in

proximity to an ester linkage.

LITERATURE CITED

(1) BEROZA, M., and GREEN, N.

1963. MATERIALS TESTED AS INSECT ATTRACTANTS.

U.S. Dept. Agr. Handb. 239, 148 pp.

(2) DETHIER, V. G.

1966. FEEDING BEHAVIOUR. 7n Haskell, P. T., ed.,

Insect Behaviour. Symposia of the Royal

Entomological Society of London, No. 3,

pp. 46-58. Bartholomew Press, Dorking,

England.

(3) BROWNE, L. B., and SMITH, C. N.

1960. THE DESIGNATION OP CHEMICALS IN TERMS

OF THE RESPONSES THEY ELICIT FROM INSECTS. Jour. Eeon. Ent. 53: 134-136.

(4) GELPERIN, A.

1966. THE CONTROL OP CROP EMPTYING IN THE

BLOW PLY. Jour. Insect Physiol. 12: 333-

347.

(5)

1966. INVESTIGATIONS OF A POREGUT RECEPTOR

ESSENTIAL TO TASTE THRESHOLD REGULATION

IN THE BLOW FLY. Jour. Insect Physiol. 12:

829-841.

(6) GREEN, N., BEROZA, M., and HALL, S. A.

1960. RECENT DEVELOPMENTS IN CHEMICAL ATTRACTANTS FOR INSECTS. In Advn. Pest Control

Res., V. Ill, pp. 129-179, Interscience,

New York.

(7) GUPTA, P. D., and THORSTEINSON, A. J.

1960. POOD PLANT RELATIONSHIPS OF THE DIAMOND-BACK MOTH (PLUTELLA MACULIPENNIS

(CURT.) >. I. GUSTATION AND OLFACTION IN

RELATION TO BOTANICAL SPECIFICITY OF THE

LARVA. Ent. Expt. et AppL 3: 241-260.

(8) JACOBSON, M.

1965. INSECT SEX ATTRACTANTS. 154 pp. John

Wiley & Sons, New York.

(9) LAAKE, E. W., PARMAN, D. C., BISHOPP, F. C, and

ROARK, R. C.

1931. THE CHEMOTROPIC RESPQNSES OP THE HOUSE

PLY, THE GREEN BOTTLE PLIES, AND THE

BLACK BLOW FLY. U.S. Dept. Agr. Tech.

Bul. 270, 10 pp.

(10) LABRECQUE, G. C., and WILSON, H. G.

1959. LABORATORY TESTS WITH SIXTY-FIVE COMPOUNDS AS REPELLENTS AGAINST HOUSE

PLIES. Pia. Ent. 42: 175-177.

(11) MAYER, M. S., and THAGGARD, C. W.

1966. INVESTIGATIONS OP AN OLFACTORY ATTRACTANT SPECIFIC FOR MALES OF THE HOUSE PLY

MUSCA DOMESTICA. JOUF. Insect Physiol.

12:891-897,

(12) PARMAN, D. C., BISHOPP, F. C., LAAKE, E. W., and

others.

1927. CHEMOTROPIC TESTS WITH THE SCREW-WORM

FLY. U.S. Dept. Agr. Bul. 1472, 32 pp.

(13) RALSTON, A. W., and BARRETT, J. P.

1941. INSECT REPELLENT ACTIVITY OP FATTY ACID

DERIVATIVES. Oil and Soap 18, pp. 89-91.

(14) RICHARDSON, C. H.

1917. THE RESPONSE OP THE HOUSE PLY (MUSCA

DOMESTICA L.) TO AMMONIA AND OTHER SUBSTANCES. N.J. Agr. Expt. Sta. Bui. 292,

19 pp.

(15) WILSON, H.G., LABRECQUE, G. C., and GAHAN, J. B.

1961. LABORATORY TESTS OF SELECTED HOUSE FLY

REPELLENTS. Fla. Ent. 44: 123-124.

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