LIB- June 1947 E-729 united States Department of Agriculture Agricultural Research Administration Bureau of Entomology and Plant Quarantine PRELIMINARY TESTS OF SYNTHETIC ORGANIC COMPOUNDS AS INSECTICIDES. PART III. By G. T. Bottger and Clemence Levin 1/ Division of Control Investigations This is the third of a series of preliminary tests on synthe- tic organic compounds being conducted at the insecticide-testing laboratory of this Bureau at Sanford, Fla. The results of the previous tests were reported in E-621 (1944) and E-634 (1945). Dur- ing 1944 a total of 139 synthetic organic compounds were tested. All the materials were tested as dusts against three or more leaf- feeding insects. Results of these dust tests served as the basis for elimination of a compound from further testing or for making subsequent tests for further evaluation of its insecticidal potenti- alities. Equipment and certain methods employed have been described by Swingle, Phillips, and Gahan. 2/ Most of the tests reported in this paper were run in triplicate, usually with 24 larvae. Compounds causing 75 percent or higher mortality of at least two species of insects were given subsequent tests. Phyto toxicity tests were made of those compounds which showed promise as possible stomach poisons. Although no attempt has been made to determine lethal doses or comparative toxicity of most of these compounds, sufficient data are presented to indicate their toxicity to insects; their utility as stomach poisons, contact insecticides, or fumigants; and their toxicity to tender foliage. Sixteen species of insects were employed in making these tests. All compounds were tested against a minimum of three species. The more toxic materials in general were tested against more species and greater numbers of insects than were the less toxic compounds. 1/ The writers wish to acknowledge the assistance of members of the Division of Insecticide Investigations, who supplied the samples for testing. Grateful acknowledgement of technical assistance is made to E. R. McGovran, formerly of the Division of Control Investigations. 2/ Swingle, M. C, Phillips, A. M., and Gahan, J. B. Laboratory testing of natural and synthetic organic substances as insecticides. Jour. Econ. Ent. 34: 95-99. 1941. JUL 2 s mi - 2 - The following insects were usedx Bean leaf roller (Urfaanos proteus (L.)) Cabbage looper (Trichoplusia ni (Hbn.)) Cotton aphid (Aphis gossypii (Glover ) Cross-striped cabbageworm (Evergestis rimosalis (Guen.)) Greenhouse leaf tier (Phlyctaenia rubigalis (Guen*)) Hawaiian beet webworm (Hymenia recurvalis (F.)) Imported cabbageworm (Pleris rapae (L.)) Large milkweed bug (Onoopeltus fasciatus (Pall.)) Melonworm (Diaphania hyalinata (L.) Pioklewoim (Diaphania nitidalis (Stoll)) Pseudoplusia looper (Pseudoplusia rogationis (Guen.)) Southern armyworm (Prodenia eridania (Cram.)) Southern beet webworm (Paohyzancla b'ipunctalis (F.) Squash bug (Anasa tristis (Peg.)) Sweetpotato weevil (Cylas formicarius elegantulus (Sum.)) Three-striped blister beetle (Epicauta lemnisoata (F.)) Eleven of the species employed as test insects were reared in the laboratory, in order to insure their availability when needed and to also provide a more or less standardized population. The Pseudoplusia looper, the bean leaf roller, the blister beetle, the cotton aphid, and the squash bug were field-collected, although comparatively few insects of any of these species were utilised. The three species most generally used (larval stage) were the southern beet webworm, the melonworm, and the southern armyworm; their susoeptibility to DPT, tested as a standard in conjunction with most tests of new samples, being in the order named. Ground derris root (4.2-4.8 percent rotenone), lead arsenate, nicotine, and pyrethrum were employed as standards for some of the tests. Most of the samples received for testing during 1944 were furnished in diluted form, usually 25 or 50 peroent in pyrophyllite, and nearly all were in satisfactory physical condition for testing. In a few instanoes, however, it was necessary to prepare the material further by grinding it in a mortar. Owing to their moisture-holding characteristic a few materials reoeived could not be dusted. - 3 - METHODS OF TESTING Dust tests. -- The first tests were made by feeding to larvae leaf sections that had been rather heavily dusted with the diluted sample material* The dust was applied in a settling chamber (de- scribed by Swingle, Phillips, and Gahan) to both surfaces of three leaves or leaf sections out to fit a petri dish, the amount of deposit being determined by weighing an aluminum plate dusted at the same time. Each leaf section was then placed in a 9-cm. petri dish, into which eight fourth-instar larvae were then introduced. After 1 and 2 days any larvae that had died were removed from the dishes. On the third day final counts of dead larvae were made, percentages of mortality were determined, and estimates made of the amount of feeding on the leaves. Spray tests. ~ Samples that showed considerable toxicity in dust tests were applied as sprays, in the proportion of 8 pounds of the oompound to 100 gallons of water. The more toxic materials were tested further at lower concentrations. Saponin was used as a wetting agent, usually in the proportion of 1/8 pound per 100 gallons of water. Leaves or plants were thoroughly sprayed on both sides and allowed to dry before being cut into sections and placed in petri dishes with test larvae. Fumigation tests « — To determine toxicity by fumigation approx- imately 1/3 gram of material was placed between two sheets of filter paper, which were then fitted into the lid of a petri dish, and eight test larvae were placed in the lower half of the dish with untreated foliage. Foliage-injury tests. — After the toxicity of a sample had been established by the petri-dish dust and spray tests, the material was applied as a spray to several truok-crop plants to determine whether it would injure tender foliage. Each compound was dissolved in water or made into a suspension spray at the rate of 8 pounds per 100 gal- lons and applied with a hand sprayer to plants growing in a garden. The plants were subjected to natural outdoor atmospheric conditions, except that they were protected from rain in order to retain as much of the residue as possible. From 6 to 8 days after being sprayed the plants were usually examined for any trace of burning or other injury resulting from the treatment. If no injury was apparent, a second application was made, and final observations for detection of injury were made at the olose of another 6-to 8-day period. Plants used for foliage-injury tests included bean, beet, collard, cotton, pea, pumpkin, Swiss chard, and tomato. - 4 - Contact tests. — Compounds showing high toxicity to certain species in dust tests were tested against one of these species in order to determine whether the material had acted as a stomach or contact poison. Additional tests were made against sucking insects to evaluate compounds as contact poisons. Volatility tests. — Weighed glass slides were dusted with the compounds and held at a constant temperature of 80 F» for 5 days, and then weighed again to determine losses due to volatilization. Volatility was determined as percentage of the original weight. DISCUSSION OF RESULTS In the majority of tests reported DDT in pyrophyllite was employed as a standard for comparing the results obtained on new compounds, although other standards were used in a few instances. Results of laboratory toxicity tests of these materials are pre- sented in table 1. The results of dust tests of all the compounds showing appre- ciable toxicity are presented in table 2. The following eight compounds were effective, killing 89 percent or more of all the leaf -feeding species on which they were tested, with only slight feeding in most cases* N-Anilino-alpha-toluamide js-Bromo-NjN-dimethylbenzene sulfonamide 2,2-Bis(pj-bromophenyl)-l-trichloroethane 1,1-Dichloro- 2, 2-bis(p_-chlorophenyl) ethane l,4-Dichloro-2-»nitrobenzene 2,2-Di-p-anisyl-l-trichloroethane alpha, aTpha'-Eexachloro-jB-xylene alpha, alpha, alpha-2, 4-Pentachlorotoluene Another 12 of the compounds were effective, killing 90 percent or more of at least 50 percent of the species, but no compound was effective against all species. Treatments of an additional five of the compounds resulted in high mortalities against one or more species, but not 50 percent of those on which they were tested. Results of various tests, including both dust and spray treat- ments, indicate that only the following four compounds were effective stomach insecticides against two or more species of leaf-feeding in- sects, and caused no more than slight injury to a small percentage of the plants tested: - 5 - p-E romo-N, N-dimethylbenzene sul f onamide 2, 2-Bis-(pj-bromophenyl)-l-tri chloroethane l,l-Dic&loro-2,2-bis(j>-chlorophenyl) ethane 2 , 2-Di-p_-ani syl-L»tri chloroethane These compounds were also effective against certain species as con- tact insecticides* However, some species which were killed as a result of feeding on treated foliage were not killed by contact treatment alone. None of these materials caused more than slight foliage injury to one or two of the five or more varieties of plants treated. Most of these compounds caused no injury at all to tender foliage, Moreover, volatility tests indicated that none of the materials were too volatile for practical use as stomach poisons* Results of fumigation tests indicate that the following compounds kill by fumigation: c 4, 6-Dinitro-m-cresyl methyl ether l,4-Dichloro-2-nitrobenzene N-Phenylglycinonitrile alpha, alpha, alpha-2,4 - Pentachlorotoluene alpha , alpha ' -Hexachlo ro-jg-xyl ene The following compounds were effective against most of the different insects when applied as contact insecticides: 4, 6-Dinitro-m-cresyl methyl ether £-Bromo-K,N-dimethylbenzenesulf onamide 4-(alpha-Thiotoluyl)morpholine l,4-Dichloro-2-nitrobenzene 2, 2-Bi s (o-bromophenyl) -1-trichloroe thane 4,6-Dinitro-o-cresyl ethyl ether 2 , 4-T ri chlo romethyl-s-tri oxane N-Pheny 1 gly cinoni t r il e 1, l-Dichloro-2, 2-bi s (£-chlorophenyl) ethane It is possible that some of the materials listed as nontoxic would show toxicity to certain insects not employed in these tests. •d 6 • o * •p C9 bO © d si o o •H ° «° © «H n 5. ■s 9 i-p o to g- © •d «o © go -P o * © a to a at © -H IS •H +> "d o © • -P «H to w *d I ! •3 E-t ©£. to«n at O Pi 0 -H © o IP-. a to pi 3 © © En O a) O En -P o © a •d s o a, o -. 6 - <«* iH C7> CO £* 00 CO o o iH 888 iH iH iH O O Q O O O iH rH iH O ■* CO o t- -> -H ® 5- E 4 & © as 5h a © 1>1 -p O 0} cl to ^. © d ■p x> ,0 0© O fe rH U Ti u •d >>rH © © ©SO •d o o< strip rn ar oaf r orm © rH iH M ■d -H 0 0 to © 1 © H g © .d 6 Qi © «S rH g| SrS ,d O © © tO A3 .M » >-. ,0 O 4> at if H in 1 a! co 0 ^ O rH a? 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